Supplemental Table 1
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Carlisle et al. 1 Supplementary Table 1. Primers for PCR amplification of bacterial 16S rRNA. Primers for Group A TA-27FMID1 CGTATCGCCTCCCTCGCGCCATCAGACGAGTGCGTAGAGTTTGATCCTGGCTCAG TA-27FMID2 CGTATCGCCTCCCTCGCGCCATCAGACGCTCGACAAGAGTTTGATCCTGGCTCAG TA-27FMID3 CGTATCGCCTCCCTCGCGCCATCAGAGACGCACTCAGAGTTTGATCCTGGCTCAG TA-27FMID4 CGTATCGCCTCCCTCGCGCCATCAGAGCACTGTAGAGAGTTTGATCCTGGCTCAG TA-27FMID5 CGTATCGCCTCCCTCGCGCCATCAGATCAGACACGAGAGTTTGATCCTGGCTCAG TA-27FMID6 CGTATCGCCTCCCTCGCGCCATCAGATATCGCGAGAGAGTTTGATCCTGGCTCAG TA-27FMID7 CGTATCGCCTCCCTCGCGCCATCAGCGTGTCTCTAAGAGTTTGATCCTGGCTCAG TA-27FMID8 CGTATCGCCTCCCTCGCGCCATCAGCTCGCGTGTCAGAGTTTGATCCTGGCTCAG TA-27FMID9 CGTATCGCCTCCCTCGCGCCATCAGTAGTATCAGCAGAGTTTGATCCTGGCTCAG TA-27FMID10 CGTATCGCCTCCCTCGCGCCATCAGTCTCTATGCGAGAGTTTGATCCTGGCTCAG TA-27FMID11 CGTATCGCCTCCCTCGCGCCATCAGTGATACGTCTAGAGTTTGATCCTGGCTCAG TA-27FMID13 CGTATCGCCTCCCTCGCGCCATCAGCATAGTAGTGAGAGTTTGATCCTGGCTCAG TA-27FMID14 CGTATCGCCTCCCTCGCGCCATCAGCGAGAGATACAGAGTTTGATCCTGGCTCAG TB-338R CTATGCGCCTTGCCAGCCCGCTCAGTGCTGCCTCCCGTAGGAGT Primers for Group B TB-27FMID1 CTATGCGCCTTGCCAGCCCGCTCAGACGAGTGCGTAGAGTTTGATCCTGGCTCAG TB-27FMID2 CTATGCGCCTTGCCAGCCCGCTCAGACGCTCGACAAGAGTTTGATCCTGGCTCAG TB-27FMID3 CTATGCGCCTTGCCAGCCCGCTCAGAGACGCACTCAGAGTTTGATCCTGGCTCAG TB-27FMID4 CTATGCGCCTTGCCAGCCCGCTCAGAGCACTGTAGAGAGTTTGATCCTGGCTCAG TB-27FMID5 CTATGCGCCTTGCCAGCCCGCTCAGATCAGACACGAGAGTTTGATCCTGGCTCAG TB-27FMID6 CTATGCGCCTTGCCAGCCCGCTCAGATATCGCGAGAGAGTTTGATCCTGGCTCAG TB-27FMID7 CTATGCGCCTTGCCAGCCCGCTCAGCGTGTCTCTAAGAGTTTGATCCTGGCTCAG TB-27FMID8 CTATGCGCCTTGCCAGCCCGCTCAGCTCGCGTGTCAGAGTTTGATCCTGGCTCAG TB-27FMID9 CTATGCGCCTTGCCAGCCCGCTCAGTAGTATCAGCAGAGTTTGATCCTGGCTCAG TB-27FMID10 CTATGCGCCTTGCCAGCCCGCTCAGTCTCTATGCGAGAGTTTGATCCTGGCTCAG TB-27FMID11 CTATGCGCCTTGCCAGCCCGCTCAGTGATACGTCTAGAGTTTGATCCTGGCTCAG TB-27FMID12 CTATGCGCCTTGCCAGCCCGCTCAGTACTGAGCTAAGAGTTTGATCCTGGCTCAG TB-27FMID13 CTATGCGCCTTGCCAGCCCGCTCAGCATAGTAGTGAGAGTTTGATCCTGGCTCAG TB-27FMID14 CTATGCGCCTTGCCAGCCCGCTCAGCGAGAGATACAGAGTTTGATCCTGGCTCAG TTA-338R CGTATCGCCTCCCTCGCGCCATCAGTGCTGCCTCCCGTAGGAGT Supplemental Table 2. Microbiota mRNA sequencing coverage. Coverage ranged from 92.8%- 100% (average 98.6%). Sample Diet Number of sequences after quality Percent Coverage trimming 1a FF 323 93 1b FF 833 98 1c FF 816 99 2a FF 662 98 2b FF 888 98 2c FF 884 99 3a FF 564 97 3b FF 966 99 4a FF 723 98 4b FF 1103 99 5a FF 746 97 10a MF 852 100 3c MF 699 100 4c MF 1154 100 5b MF 1144 100 6a MF 963 100 6b MF 919 100 7a MF 888 99 7b MF 1096 100 8a MF 834 100 9a MF 961 99 Carlisle et al. 2 Supplemental Table 3. Library characteristics for 8 cDNA libraries. Average read length was 100bp. BLAST search against the mouse RNA database yielded 18,014,657 hits (~2,251,000/library) to 19,703 mouse mRNAs. FF1 FF2 FF3 FF4 MF1 MF2 MF3 MF4 Total reads before quality trim 4,096,796 6,609,830 3,579,743 3,452,378 2,580,264 3,781,577 3,320,574 2,075,326 %GC 62 62 62 62 63 61 62 61 Reads for annotation 3,902,252 6,304,521 3,523,485 3,397,021 2,460,776 3,601,183 3,269,729 2,042,846 Total Reads Annotated 1,801,239 5,309,548 2,558,030 2,038,522 785,627 2,087,931 2,384,179 1,049,581 Mouse Genes Detected 11,695 16,273 13,166 13,261 9,943 13,191 13,141 11,921 Carlisle et al. 3 Supplemental Table 4. The 50 genes with the highest mRNA transcript levels. mRNA transcript level of each gene was calculated for FF and MF mice. p<0.01 indicated a statistically significant difference. Gene p-value Gene p-value 28S rRNA (LOC236598) Rn28s 0.10 Keratin 8 Krt8 0.58 18S rRNA Rn18s 0.19 RNA component of mitochondrial RNAse P RMrp 0.43 Leucyl-tRNA synthetase Lars2 0.89 Small praline-rich protein 2A2 Sprr2a 0.47 mitocohondrial nuclear gene 2 encoding mitochondrial protein microRNA 689-2 Mir689-2 0.04 Ribosomal protein, large P0 Rplp0 0.89 Ac1147 (LOC672857) Ac1147 0.73 7SK nuclear Rn7sk 0.07 MicroRNA mir-2135-3 Mir 21353 0.89 Metallothionein 2 Mt2 0.01 Transmembrane protein 170 Tmem170 0.82 Lectin, galactose binding, soulable 4 Lgals4 0.74 LOC627695 0.90 Actin, alpha 2, smooth muscle Acta2 0.30 Ribonuclease P RNA component Rpph1 0.81 Actin, gamma, cytoplasmic 1 Actg1 0.30 H1 microRNA mir 2133-2 Mir2133- 0.30 Tumor protein, translationally-controlled 1 Tpt1 0.81 2 Histone 3 (LOC677333) H3 0.32 Collagen, type 1, alpha 2 Col1a2 0.11 Eukaryotic translation elongation Eef1a1 0.50 Keratin 19 Krt19 0.82 factor 1 alpha 1 Collagen type III alpha 1 Col3a1 0.14 Filamin, alpha Flna 0.15 Mtrnr2-like Mtrnr2l 0.76 Actin, gamma 2, smooth muscle, enteric Actg2 0.04 Actin, beta Actb 0.06 Ferritin heavy chain 1 Fth1 0.05 Metallothionein 1 Mt1 0.003 Glyceraldehyde-3-phosphate dehydrogenase Gapdh 0.58 Collagen, type 1, alpha 1 Col1a1 0.09 Heat shock protein 8 Hspa8 0.53 Proline/histidine/glycine-rich 1 Phgr1 0.69 microRNA mir-2134-4 Mir213 0.11 4-4 Deleted in malignant brain tumors Dmbt1 0.800 Transgelin Tagln 0.45 1 Mucin 2 Muc2 0.975 Ribosomal protein SA Rpsa 0.75 Fc fragment of IgG binding protein Fcgbp 0.802 Serine protease inhibitor, Kunitz type 2 Spint2 0.91 5.8S ribosomal RNA 5.8S 0.009 Myosin, heavy polypeptide 11, smooth muscle Myh11 0.36 rRNA RIKEN cDNA 2210415F13 0.347 ATPase, Na/K transporting, alpha 1 Atp1a1 0.36 polypeptide Ferritin light chain 1 Flt1 0.180 ATP synthase, H+transporting mitochondrial Atp5b 0.36 F1 complex, beta subunit Eukaryotic translation elongation Eef2 0.944 Ubiquitin B Ubb 0.64 factor 2 Carlisle et al. 4 Supplemental Table 5. Host transcriptome analysis. mRNA transcript levels differed between MF and FF mice for 148 genes. p<0.01 was considered statistically significant. Fold change in FF mice relative to MF mice was calculated for each gene. Literature search for each gene was conducted, and genes were grouped based on published function: C/G-Cell Cycle/Gene Expression (n=32), SI-Structural Integrity (n=6), SR-Stress Response (n=18), NU-Nutrient Utilization (n=6), IR-Immune Regulation (n=14), CM-Cell Motility (n=6), VF-Vascular Function (n=3)). Gene Fold p-value Function Gene Fold p-value Function Change Change Methallothionein 1 Mt1 5.40↑ 0.0030 Autophagy-related 4B Atg4b 0.37↓ 0.0028 SR25, 26 5.8S ribosomal RNA 5.8S rRNA 1.7↑ 0.0095 Pirin Pir FF only 0.0022 C/G46 Desmin Des 0.56↓ 0.0016 SI32 Engulfment and cell motility 2, ced- Elmo2 FF only 0.0038 SR47 12 homolog Cysteine and glycine-rich Csrp1 0.66↓ 0.0065 Sestrin 3 Sesn3 0.41↓ 0.0040 SR27 protein 1 Tropomyosin 2 beta Tpm2 0.63↓ 0.0068 Regulator of G-protein signaling 3 Rgs3 0.43↓ 0.0097 IR48 Heme Oxygenase 1 Hmox1 25.4↑ 0.0066 SR 17, 18 Protein inhibitor of activated STAT2 Pias2 FF only 0.0048 C/G49 Serine pepditase inhibitor Serpinh1 0.49↓ 0.0074 SI31 Autophagy-related 5 Atg5 0.33↓ 0.0052 SR25 Cladue H member 1 ATP-binding cassette, sub- Abcbia 2.68↑ 0.0075 SR23 Kinesin family member 22 Kif22 9.46↑ 0.0064 family B, member 1A Supression of turmoigenicity St14 1.94↑ 0.0003 SR50 RIKEN 9030625A04 4.86↑ 0.0072 14 Vinculin Vcl 0.47↓ 0.0061 SI29 RIKEN 9130024F11 0.37↓ 0.0082 Protein Phosphatase 1, Ppp1cb 0.55↓ 0.0057 RAN binding protein 6 RANbp FF only 0.0015 CM51 catalytic subunit, beta isoform 6 Protein tyrosine phosphatase Ptp4a1 0.67↓ 0.0083 RIKEN 9530082P21 0.32↓ 0.0046 4a1 Ribophorin 1 Rpn1 0.70↓ 0.0069 RNA binding motif protein 12 Rbm12 0.33↓ 0.0032 C/G52 Glutamate-cysteine ligase, Gclc 5.66↑ 0.0051 SR18 bolA-like 2 Bola2 FF only 0.0060 SR53 catalytic subunit Transmembrane protein 30A Tmem30a 0.58↓ 0.0019 Testis-specific kinase 2 Tesk2 ↓0.42 0.0016 Aldehyde dehydrogenase 9, Aldh9a1 1.67↑ 0.0039 ERGIC and golgi 2 Ergic2 ↓0.36 0.0029 CM54 subfamily A1 Melanoma inhibitory activity 3 Mia3 2.66↑ 0.0062 DnaJ (Hsp40) homolog, subfamily C, Dnajc21 8.61↑ 0.0057 SR21 member 21 Vacuolar protein sorting 13B Vps13b 2.97↑ 0.0036 Carbohydrate sulfotransferase 3 Chst3 FF only 0.0010 3’-phosphoadenosine 5’- Papss1 0.61↓ 0.0050 JNK1/MAPK8-associated membrane Jkamp 8.2↑ 0.0025 SR24 phosphosulfate synthase 1 protein High mobility group Hmgn2 0.39↓ 0.0073 Hypothetical protein LOC100233175 7.18↑ 0.0036 nucleosomal binding domain 2 Integrin alpha 5 Itga5 0.34↓ 0.0094 SI55, 56 COX11 homolog, cytochrome c Cox11 0.38↓ 0.0069 SR57 oxidase assembly protein Proliferating cell nuclear PCNA 0.43↓ 0.0003 C/G58 H1 histone family, member X H1fx 0.38↓ 0.0059 antigen Family with sequence Fam109a 2.82↑ 0.0073 Actin, alpha 1, skeletal muscle Acta1 0.16↓ 0.0026 similarity 109, member A Cathepsin L Ctsl 0.40↓ 0.0075 IR59 Mitochondrial translational release Mtrf1 FF only 0.0044 factor 1 Cytochrome P450, family 4, Cyp4f16 0.47↓ 0.0061 IR60 Similar to Ornithine decarboxylase ODC FF only 0.0070 subunit f, polypeptiede 16 Ribosomal protein L3 0.61↓ 0.0044 Secretogranin III Scg3 FF only 0.0021 Lymphocyte specific 1, Lsp1 0.38↓ 0.0044 IR34 SRY-box containing gene 17 Sox17 FF only 0.0074 VF61 transcript variant 2 Solute carrier family 38, Slc38a10 0.60↓ 0.0029 CM62 Phosphodiesterase 6D, cGMP- Pde6d 0.30↓ 0.0034 member 10 specific, rod, delta Metastasis suppressor 1-like Mtss1l 0.48↓ 0.0040 Hexosaminidase Hexdc FF only 0.0006 Regulation of nuclear pre- Rprd2 0.60↓ 0.0083 Similar to Chlorine interacellular FF only 0.0040 mRNA domain containing 2 channel 4 Zinc finger, CCHC domain Zcchc2 2.69↑ 0.0069 Hemopexin Hpx FF only 0.0040 SR63, IR64, containing 2 65 GTPase activating protein and Gapvd1 0.38↓ 0.0028 Cyclin T1 Ccnt1 0.25↓ 0.0094 IR66, C/G67 VPS9 domain 1 Eph receptor A2 Epha2 2.60↑ 0.0052 CM68 Mab-21 like 1 Mab21l FF only 0.0054 1 Similar to mKIAA1397 protein 3.49↑ 0.0078 Adenylosuccinate synthetase like 1 Adssl1 FF only 0.0054 NU69 Solute carrier family 35, Scl35e3 3.41↑ 0.0022 BC068157 5.00↑ 0.0090 member E3 SYF2 homolog,