Fig. 7

PRKCSH mitigates ER stress-induced cell death. a Cell death analysis (left) in L02-Mock and L02-PRK cells treated with 10 μg/mL TM or 1 μM TG for 72 h. Cell death was determined from PI staining followed by FACS analysis. Immunoblot analysis of PARP1 cleavage (right) in these cells. b Cell death analysis (left) in PRKCSH-silenced Huh-7 cells treated with 10 μg/mL TM for 48 h. Immunoblot analysis of PARP1 cleavage (right) in these cells. c Cell death analysis (left) in PRKCSH-silenced Huh-7 cells treated with 10 μg/mL TM or 1 μM TG for 72 h. Immunoblot analysis of PARP1 cleavage (right) in these cells. d Cell death analysis (left) in Huh-WT and Huh-PRK KO cells treated with 10 μg/mL TM for 72 h. Immunoblot analysis of PARP1 cleavage (right) in these cells. e Cell survival analysis in L02-Mock and L02-PRK cells (left), and Huh-WT and Huh-PRK KO cells (right). Cells were exposed to 10 μg/mL TM or EBSS (for nutrient starvation) for 48 h, and then replated in normal cell culture medium. After 5 days, cell viability was determined by staining with crystal violet. Number of cells per area (mm²) was calculated. Scale bars represent 100 µm. f In vivo tumor growth analysis in xenograft nude mice injected with Huh-PRK KO or WT cells. Balb/c nude mice were subcutaneous transplantated with each tumor cells in the right groin (four mice in each group). The volume of tumors was monitored for 35 days. Representative images of tumor-bearing mice (top) and tumor volumes (bottom) were presented. Scale bars represent 10 mm. g Cell death analysis in L02 cells transfected with the control, WT PRKCSH, ΔE/P, or MRH mutant plasmids; cells were treated with 10 μg/mL TM for 72 h. h Cell death analysis (left) in IRE1α-silenced L02-PRK cells treated with 10 μg/mL TM for 72 h. Immunoblot analysis of PARP1 cleavage (right) in these cells. In cell death analysis and cell survival analysis, data are shown as mean ± SEM from three or four independent experiments. One-way ANOVA; *P < 0.05, **P < 0.01