Supplementary Materials s29

Supplementary Materials s29

<p>Supplementary Materials Figure S1 shows additional analysis of 3-MA, SB203580 on selenite-induced apoptosis. Figure S2 shows additional data on the immunofluorescent co-localization of p38 and p53. Figure S3 shows the images of gel electrophoresis and measurements of optical density measurements of extracted RNA. </p><p>1 Figure S1 Cells were pretreated with SB203580, 3-MA and selenite as described in the text, and then cell apoptotic rates were analyzed by Annexin-V assay. Data are presented as the means±SD (n=3). *p<0.01 compared with the control group.</p><p>**p<0.05 compared with the selenite treatment group. ***p<0.01 compared with the combination treatment group.</p><p>2 Figure S2 Immunofluorescent co-localization of p38 and p53. The cells were treated with 20 μM selenite for 24 h and analyzed by immunofluorescent microscopy using antibodies against p38 (red) and p53 (green). The scale bar represents 100 μm.</p><p>3 Figure S3 Extracted RNA quality control was performed using gel electrophoresis and optical density measurements. (a-c) Cells were pretreated with SB203580, or transfected with p38-siRNA or plasmid-p38DN before selenite exposure for 24 h (a), cells were pretreated with Salubrinal, Mknk1 inhibitor, or transfected with eIF2a- siRNA or plasmid-eIF4E before selenite exposure for 24 h (b), cells were transfected with plasmid-p53MT before selenite treatment (c), and then the denaturing gel electrophoresis was conducted to visually assess the quality of extracted RNAs. </p><p>4 Supplementary table 1: siRNAs used.</p><p>Gene Targeting sequence p38 MAPK 5’- GGAAUU CAA UGA UGU GUA U -3’ Hsp90 5′-GAU CAG ACA GAG UAC CUA G-3′ PERK 5’- GCG GCA GGU CAU UAG UAA U -3’ eIF2α 5’-GUA AGG AUG GGA CAU UGU U -3’ scrambled siRNA 5'- UUC UCC GAA CGA ACG UGU CAC GU -3'</p><p>5 Supplementary table 2: Primer sequence for Real-Time PCR.</p><p>Gene Primers Atf4 Forward 5′- AAC AAC AGC AAG GAG GAT GC -3′</p><p>Reverse 5′- GCA TGG TTT CCA GGT CAT CT -3′ chop Forward 5′- TGC CTT TCA CCT TGG AGA CG -3′</p><p>Reverse 5′- CCA TAG AAC TCT GAC TGG AAT CTG G -3′</p><p> gapdh Forward 5′- AAC ATC AAA TGG GGT GAG GCC -3′</p><p>Reverse 5′- GTT GTC ATG GAT GAC CTT GGC -3</p><p>β-actin Forward 5′- CTA CAA TGA GCT GCG TGT GG -3′</p><p>Reverse 5′- AAG GAA GGC TGG AAG AGT GC -3</p><p>6 Supplementary table 3: q-PCR primers used to detect promoter enrichment by the ChIP assay.</p><p>Gene Primers Location (nt) map1lc3b-A site Forward 5′- GGA GGG GAA AGG ATG GTC GG -3′ -528 to -340</p><p>Reverse 5′- CCT GAG GTG ACG GTT GTG GG -3′ map1lc3b-a site Forward 5′- CGG TTT CAA GCG ATT CTC -3′ -1895 to -1723</p><p>Reverse 5′- ACT TTG GGA GGT CAA GGC -3′ chop-B site Forward 5′- GTG AGG GCT CTG GGA GGT GCT -3′ -875 to -689</p><p>Reverse 5′- AGG GGG ATG TTA CCT TTC CCT TCT C -3′ chop-b site Forward 5′- GGG CCC CTT CCA AAG CCA CAG T -3′ -1833 to -1564</p><p>Reverse 5′- TGG TCC ACC CTC CAC ACT ACC CCC A -3′</p><p>7</p>

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