Supplementary Materials s29

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.

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<0.05 compared with the selenite treatment group. ***p<0.01 compared with the combination treatment group.

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.

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.

4 Supplementary table 1: siRNAs used.

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'

5 Supplementary table 2: Primer sequence for Real-Time PCR.

Gene Primers Atf4 Forward 5′- AAC AAC AGC AAG GAG GAT GC -3′

Reverse 5′- GCA TGG TTT CCA GGT CAT CT -3′ chop Forward 5′- TGC CTT TCA CCT TGG AGA CG -3′

Reverse 5′- CCA TAG AAC TCT GAC TGG AAT CTG G -3′

gapdh Forward 5′- AAC ATC AAA TGG GGT GAG GCC -3′

Reverse 5′- GTT GTC ATG GAT GAC CTT GGC -3

β-actin Forward 5′- CTA CAA TGA GCT GCG TGT GG -3′

Reverse 5′- AAG GAA GGC TGG AAG AGT GC -3

6 Supplementary table 3: q-PCR primers used to detect promoter enrichment by the ChIP assay.

Gene Primers Location (nt) map1lc3b-A site Forward 5′- GGA GGG GAA AGG ATG GTC GG -3′ -528 to -340

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

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

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

Reverse 5′- TGG TCC ACC CTC CAC ACT ACC CCC A -3′