A Commercial Antibody to the Human Condensin II Subunit NCAPH2 Cross-Reacts with a SWI/SNF Complex Component
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Active Motif Technical Data Sheet (TDS)
NCAPH2 antibody (mAb) Catalog Nos: 61681, 61682 Quantities: 100 µg, 10 µg RRID: AB_2793733 Purification: Protein G Chromatography Clone: 5F2G4 Host: Rat Isotype: IgG2a Concentration: 1 µg/µl Application(s): ICC, IF, WB Molecular Weight: 90 kDa Reactivity: Human Background: NCAPH2 (Non-SMC Condensin II Complex, Subunit H2) Regulatory subunit of the condensin-2 complex, a complex that seems to provide chromosomes with an additional level of organization and rigidity and in establishing mitotic chromosome architecture. May play a role in lineage-specific role in T-cell development. (By similarity) Immunogen: This antibody was raised against a recombinant protein to human NCAPH2. Buffer: Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic. NCAPH2 antibody (mAb) (Clone 5F2G4) Application Notes: tested by immunofluorescence. Formaldehyde fixed HeLa cell stained with the Applications Validated by Active Motif: NCAPH2 antibody. ICC/IF: 1 - 2 µg/ml dilution WB*: 0.5 - 2 µg/ml dilution The addition of 0.1% Tween 20 in the blocking buffer and primary antibody incubation buffer is recommended to aid in detection by Western blot. Individual optimization may be required. *Note: many chromatin-bound proteins are not soluble in a low salt nuclear extract and fractionate to the pellet. Therefore, we recommend a High Salt / Sonication Protocol when preparing nuclear extracts for Western blot. Storage and Guarantee: Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. -
NCAPD3 Antibody (C-Term) Affinity Purified Rabbit Polyclonal Antibody (Pab) Catalog # AP16786B
10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 NCAPD3 Antibody (C-term) Affinity Purified Rabbit Polyclonal Antibody (Pab) Catalog # AP16786B Specification NCAPD3 Antibody (C-term) - Product Information Application WB,E Primary Accession P42695 Other Accession NP_056076.1 Reactivity Human Host Rabbit Clonality Polyclonal Isotype Rabbit Ig Calculated MW 168891 Antigen Region 1050-1078 NCAPD3 Antibody (C-term) - Additional Information NCAPD3 Antibody (C-term) (Cat. Gene ID 23310 #AP16786b) western blot analysis in K562 cell line lysates (35ug/lane).This Other Names Condensin-2 complex subunit D3, Non-SMC demonstrates the NCAPD3 antibody detected condensin II complex subunit D3, hCAP-D3, the NCAPD3 protein (arrow). NCAPD3, CAPD3, KIAA0056 Target/Specificity NCAPD3 Antibody (C-term) - Background This NCAPD3 antibody is generated from rabbits immunized with a KLH conjugated Condensin complexes I and II play essential synthetic peptide between 1050-1078 roles in amino acids from the C-terminal region of mitotic chromosome assembly and human NCAPD3. segregation. Both condensins contain 2 invariant structural maintenance of Dilution chromosome (SMC) WB~~1:1000 subunits, SMC2 (MIM 605576) and SMC4 (MIM 605575), but they contain Format different sets of non-SMC subunits. NCAPD3 is Purified polyclonal antibody supplied in PBS 1 of 3 non-SMC with 0.09% (W/V) sodium azide. This subunits that define condensin II (Ono et al., antibody is purified through a protein A 2003 [PubMed column, followed by peptide affinity 14532007]). purification. NCAPD3 Antibody (C-term) - References Storage Maintain refrigerated at 2-8°C for up to 2 Rose, J.E., et al. -
A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated. -
Condensation of Prometaphase Chromo-Somes
Condensation of Prometaphase Chromo- somes D'Eustachio, P., Matthews, L., Orlic-Milacic, M. European Bioinformatics Institute, New York University Langone Medical Center, Ontario Institute for Cancer Research, Oregon Health and Science University. The contents of this document may be freely copied and distributed in any media, provided the authors, plus the institutions, are credited, as stated under the terms of Creative Commons Attribution 4.0 Inter- national (CC BY 4.0) License. For more information see our license. 09/03/2019 Introduction Reactome is open-source, open access, manually curated and peer-reviewed pathway database. Pathway annotations are authored by expert biologists, in collaboration with Reactome editorial staff and cross- referenced to many bioinformatics databases. A system of evidence tracking ensures that all assertions are backed up by the primary literature. Reactome is used by clinicians, geneticists, genomics research- ers, and molecular biologists to interpret the results of high-throughput experimental studies, by bioin- formaticians seeking to develop novel algorithms for mining knowledge from genomic studies, and by systems biologists building predictive models of normal and disease variant pathways. The development of Reactome is supported by grants from the US National Institutes of Health (P41 HG003751), University of Toronto (CFREF Medicine by Design), European Union (EU STRP, EMI-CD), and the European Molecular Biology Laboratory (EBI Industry program). Literature references Fabregat, A., Sidiropoulos, K., Viteri, G., Forner, O., Marin-Garcia, P., Arnau, V. et al. (2017). Reactome pathway ana- lysis: a high-performance in-memory approach. BMC bioinformatics, 18, 142. ↗ Sidiropoulos, K., Viteri, G., Sevilla, C., Jupe, S., Webber, M., Orlic-Milacic, M. -
Supplementary File 2A Revised
Supplementary file 2A. Differentially expressed genes in aldosteronomas compared to all other samples, ranked according to statistical significance. Missing values were not allowed in aldosteronomas, but to a maximum of five in the other samples. Acc UGCluster Name Symbol log Fold Change P - Value Adj. P-Value B R99527 Hs.8162 Hypothetical protein MGC39372 MGC39372 2,17 6,3E-09 5,1E-05 10,2 AA398335 Hs.10414 Kelch domain containing 8A KLHDC8A 2,26 1,2E-08 5,1E-05 9,56 AA441933 Hs.519075 Leiomodin 1 (smooth muscle) LMOD1 2,33 1,3E-08 5,1E-05 9,54 AA630120 Hs.78781 Vascular endothelial growth factor B VEGFB 1,24 1,1E-07 2,9E-04 7,59 R07846 Data not found 3,71 1,2E-07 2,9E-04 7,49 W92795 Hs.434386 Hypothetical protein LOC201229 LOC201229 1,55 2,0E-07 4,0E-04 7,03 AA454564 Hs.323396 Family with sequence similarity 54, member B FAM54B 1,25 3,0E-07 5,2E-04 6,65 AA775249 Hs.513633 G protein-coupled receptor 56 GPR56 -1,63 4,3E-07 6,4E-04 6,33 AA012822 Hs.713814 Oxysterol bining protein OSBP 1,35 5,3E-07 7,1E-04 6,14 R45592 Hs.655271 Regulating synaptic membrane exocytosis 2 RIMS2 2,51 5,9E-07 7,1E-04 6,04 AA282936 Hs.240 M-phase phosphoprotein 1 MPHOSPH -1,40 8,1E-07 8,9E-04 5,74 N34945 Hs.234898 Acetyl-Coenzyme A carboxylase beta ACACB 0,87 9,7E-07 9,8E-04 5,58 R07322 Hs.464137 Acyl-Coenzyme A oxidase 1, palmitoyl ACOX1 0,82 1,3E-06 1,2E-03 5,35 R77144 Hs.488835 Transmembrane protein 120A TMEM120A 1,55 1,7E-06 1,4E-03 5,07 H68542 Hs.420009 Transcribed locus 1,07 1,7E-06 1,4E-03 5,06 AA410184 Hs.696454 PBX/knotted 1 homeobox 2 PKNOX2 1,78 2,0E-06 -
Ncapd3 CRISPR/Cas9 KO Plasmid (M): Sc-429692
SANTA CRUZ BIOTECHNOLOGY, INC. Ncapd3 CRISPR/Cas9 KO Plasmid (m): sc-429692 BACKGROUND APPLICATIONS The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and Ncapd3 CRISPR/Cas9 KO Plasmid (m) is recommended for the disruption of CRISPR-associated protein (Cas9) system is an adaptive immune response gene expression in mouse cells. defense mechanism used by archea and bacteria for the degradation of foreign genetic material (4,6). This mechanism can be repurposed for other 20 nt non-coding RNA sequence: guides Cas9 functions, including genomic engineering for mammalian systems, such as to a specific target location in the genomic DNA gene knockout (KO) (1,2,3,5). CRISPR/Cas9 KO Plasmid products enable the U6 promoter: drives gRNA scaffold: helps Cas9 identification and cleavage of specific genes by utilizing guide RNA (gRNA) expression of gRNA bind to target DNA sequences derived from the Genome-scale CRISPR Knock-Out (GeCKO) v2 library developed in the Zhang Laboratory at the Broad Institute (3,5). Termination signal Green Fluorescent Protein: to visually REFERENCES verify transfection CRISPR/Cas9 Knockout Plasmid CBh (chicken β-Actin 1. Cong, L., et al. 2013. Multiplex genome engineering using CRISPR/Cas hybrid) promoter: drives systems. Science 339: 819-823. 2A peptide: expression of Cas9 allows production of both Cas9 and GFP from the 2. Mali, P., et al. 2013. RNA-guided human genome engineering via Cas9. same CBh promoter Science 339: 823-826. Nuclear localization signal 3. Ran, F.A., et al. 2013. Genome engineering using the CRISPR-Cas9 system. Nuclear localization signal SpCas9 ribonuclease Nat. Protoc. 8: 2281-2308. -
Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase -
Appendix 2. Significantly Differentially Regulated Genes in Term Compared with Second Trimester Amniotic Fluid Supernatant
Appendix 2. Significantly Differentially Regulated Genes in Term Compared With Second Trimester Amniotic Fluid Supernatant Fold Change in term vs second trimester Amniotic Affymetrix Duplicate Fluid Probe ID probes Symbol Entrez Gene Name 1019.9 217059_at D MUC7 mucin 7, secreted 424.5 211735_x_at D SFTPC surfactant protein C 416.2 206835_at STATH statherin 363.4 214387_x_at D SFTPC surfactant protein C 295.5 205982_x_at D SFTPC surfactant protein C 288.7 1553454_at RPTN repetin solute carrier family 34 (sodium 251.3 204124_at SLC34A2 phosphate), member 2 238.9 206786_at HTN3 histatin 3 161.5 220191_at GKN1 gastrokine 1 152.7 223678_s_at D SFTPA2 surfactant protein A2 130.9 207430_s_at D MSMB microseminoprotein, beta- 99.0 214199_at SFTPD surfactant protein D major histocompatibility complex, class II, 96.5 210982_s_at D HLA-DRA DR alpha 96.5 221133_s_at D CLDN18 claudin 18 94.4 238222_at GKN2 gastrokine 2 93.7 1557961_s_at D LOC100127983 uncharacterized LOC100127983 93.1 229584_at LRRK2 leucine-rich repeat kinase 2 HOXD cluster antisense RNA 1 (non- 88.6 242042_s_at D HOXD-AS1 protein coding) 86.0 205569_at LAMP3 lysosomal-associated membrane protein 3 85.4 232698_at BPIFB2 BPI fold containing family B, member 2 84.4 205979_at SCGB2A1 secretoglobin, family 2A, member 1 84.3 230469_at RTKN2 rhotekin 2 82.2 204130_at HSD11B2 hydroxysteroid (11-beta) dehydrogenase 2 81.9 222242_s_at KLK5 kallikrein-related peptidase 5 77.0 237281_at AKAP14 A kinase (PRKA) anchor protein 14 76.7 1553602_at MUCL1 mucin-like 1 76.3 216359_at D MUC7 mucin 7, -
Interactome Analyses Revealed That the U1 Snrnp Machinery Overlaps Extensively with the RNAP II Machinery and Contains Multiple
www.nature.com/scientificreports OPEN Interactome analyses revealed that the U1 snRNP machinery overlaps extensively with the RNAP II Received: 12 April 2018 Accepted: 24 May 2018 machinery and contains multiple Published: xx xx xxxx ALS/SMA-causative proteins Binkai Chi1, Jeremy D. O’Connell1,2, Tomohiro Yamazaki1, Jaya Gangopadhyay1, Steven P. Gygi1 & Robin Reed1 Mutations in multiple RNA/DNA binding proteins cause Amyotrophic Lateral Sclerosis (ALS). Included among these are the three members of the FET family (FUS, EWSR1 and TAF15) and the structurally similar MATR3. Here, we characterized the interactomes of these four proteins, revealing that they largely have unique interactors, but share in common an association with U1 snRNP. The latter observation led us to analyze the interactome of the U1 snRNP machinery. Surprisingly, this analysis revealed the interactome contains ~220 components, and of these, >200 are shared with the RNA polymerase II (RNAP II) machinery. Among the shared components are multiple ALS and Spinal muscular Atrophy (SMA)-causative proteins and numerous discrete complexes, including the SMN complex, transcription factor complexes, and RNA processing complexes. Together, our data indicate that the RNAP II/U1 snRNP machinery functions in a wide variety of molecular pathways, and these pathways are candidates for playing roles in ALS/SMA pathogenesis. Te neurodegenerative disease Amyotrophic Lateral Sclerosis (ALS) has no known treatment, and elucidation of disease mechanisms is urgently needed. Tis problem has been especially daunting, as mutations in greater than 30 genes are ALS-causative, and these genes function in numerous cellular pathways1. Tese include mitophagy, autophagy, cytoskeletal dynamics, vesicle transport, DNA damage repair, RNA dysfunction, apoptosis, and pro- tein aggregation2–6. -
Non-SMC Condensin I Complex Subunit H Mediates Mature
www.nature.com/scientificreports OPEN Non-SMC condensin I complex subunit H mediates mature chromosome condensation and DNA damage in pancreatic cancer cells Jae Hyeong Kim 1*, Yuna Youn1, Kyung-Tae Kim2, Gyubeom Jang2 & Jin-Hyeok Hwang1,3* Non-SMC condensin I complex subunit H (NCAPH) is a vital gene associated with chromosome stability and is required for proper chromosome condensation and segregation. However, the mechanisms through which NCAPH afects pancreatic cancer (PC) and its molecular function remain unclear. In this study, we examined the role of NCAPH in PC cells. Our results showed that NCAPH was overexpressed in clinical PC specimens (GEPIA) and cell lines. In addition, in NCAPH-knockdown cells, colony formation and proliferation were inhibited, and the cell cycle was arrested at the S and G2/M phases owing to failure of mature chromosome condensation (MCC) in poorly condensed chromosomes. Increased cell death in NCAPH-knockdown cells was found to help initiate apoptosis through the activation of caspase-3 and PARP cleavage. Furthermore, NCAPH-knockdown cells showed an increase in chromosomal aberrations and DNA damage via activation of the DNA damage response (Chk1/Chk2) signaling pathways. These data demonstrated that NCAPH played an important role in cell cycle progression and DNA damage by maintaining chromosomal stability through progression of MCC from poorly condensed chromosomes. Ultimately, NCAPH knockdown induced apoptotic cell death, which was partially mediated by caspase-dependent pathways. These fndings highlight the potential role of NCAPH as a therapeutic target for PC. More than 90% of pancreatic cancer (PC) cases are classifed as pancreatic ductal adenocarcinoma (PDA), one of the most lethal human malignant tumors with a poor prognosis1. -
A Yeast Phenomic Model for the Influence of Warburg Metabolism on Genetic Buffering of Doxorubicin Sean M
Santos and Hartman Cancer & Metabolism (2019) 7:9 https://doi.org/10.1186/s40170-019-0201-3 RESEARCH Open Access A yeast phenomic model for the influence of Warburg metabolism on genetic buffering of doxorubicin Sean M. Santos and John L. Hartman IV* Abstract Background: The influence of the Warburg phenomenon on chemotherapy response is unknown. Saccharomyces cerevisiae mimics the Warburg effect, repressing respiration in the presence of adequate glucose. Yeast phenomic experiments were conducted to assess potential influences of Warburg metabolism on gene-drug interaction underlying the cellular response to doxorubicin. Homologous genes from yeast phenomic and cancer pharmacogenomics data were analyzed to infer evolutionary conservation of gene-drug interaction and predict therapeutic relevance. Methods: Cell proliferation phenotypes (CPPs) of the yeast gene knockout/knockdown library were measured by quantitative high-throughput cell array phenotyping (Q-HTCP), treating with escalating doxorubicin concentrations under conditions of respiratory or glycolytic metabolism. Doxorubicin-gene interaction was quantified by departure of CPPs observed for the doxorubicin-treated mutant strain from that expected based on an interaction model. Recursive expectation-maximization clustering (REMc) and Gene Ontology (GO)-based analyses of interactions identified functional biological modules that differentially buffer or promote doxorubicin cytotoxicity with respect to Warburg metabolism. Yeast phenomic and cancer pharmacogenomics data were integrated to predict differential gene expression causally influencing doxorubicin anti-tumor efficacy. Results: Yeast compromised for genes functioning in chromatin organization, and several other cellular processes are more resistant to doxorubicin under glycolytic conditions. Thus, the Warburg transition appears to alleviate requirements for cellular functions that buffer doxorubicin cytotoxicity in a respiratory context. -
Supplementary Table 1
Supplementary Table 1. Phosphoryl I-Area II-Area II-Debunker III-Area Gene Symbol Protein Name Phosphorylated Peptide I-R2 I-Debunker score II-R2 III-R2 ation Site Ratio Ratio score Ratio 92154 ABBA-1 Actin-bundling protein with BAIAP2 homologyK.TPTVPDS*PGYMGPTR.A S601 1.76 0.95 0.999958250 1.18 0.98 0.999971836 0.98 0.98 92154 ABBA-1 Actin-bundling protein with BAIAP2 homologyR.AGS*EECVFYTDETASPLAPDLAK.A S612 1.40 0.99 0.999977464 1.03 0.99 0.999986373 1.00 0.99 92154 ABBA-1 Actin-bundling protein with BAIAP2 homologyK.GGGAPWPGGAQTYS*PSSTCR.Y S300 0.49 0.98 0.999985406 0.97 0.99 0.999983906 2.03 0.97 23 ABCF1 ATP-binding cassette sub-family F memberK.QQPPEPEWIGDGESTS*PSDK.V 1 S22 1.09 0.98 0.872361494 0.81 1.00 0.847115585 0.97 0.97 27 ABL2 Isoform IA of Tyrosine-protein kinase ABL2K.VPVLIS*PTLK.H S936 0.76 0.98 0.999991559 1.06 0.99 0.999989547 0.99 0.96 3983 ABLIM1 Actin binding LIM protein 1 R.TLS*PTPSAEGYQDVR.D S433 1.27 0.99 0.999994010 1.16 0.98 0.999989861 1.11 0.99 31 ACACA acetyl-Coenzyme A carboxylase alpha isoformR.FIIGSVSEDNS*EDEISNLVK.L 1 S29 1.23 0.99 0.999992898 0.72 0.99 0.999992499 0.83 0.99 65057 ACD Adrenocortical dysplasia protein homologR.TPS*SPLQSCTPSLSPR.S S424 1.51 0.90 0.999936226 0.82 0.88 0.997623142 0.46 0.93 22985 ACIN1 Apoptotic chromatin condensation inducerK.ASLVALPEQTASEEET*PPPLLTK.E in the nucleus T414 0.90 0.92 0.922696554 0.91 0.92 0.993049132 0.95 0.99 22985 ACIN1 Apoptotic chromatin condensation inducerK.ASLVALPEQTAS*EEETPPPLLTK.E in the nucleus S410 1.02 0.99 0.997470008 0.80 0.99 0.999702808 0.96