Fig. 4

LINP1 functions in repressing UPR and downstream apoptotic genes. A, C qPCR validations of key gene expression in endoplasmic reticulum signaling including GRP78, XBP1, DDIT3 and DR5 in response to LINP1 knockout and overexpression. Each experiment was performed in at least triplicate and results are presented as mean ± s.d. One-way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (*P < 0.05, **P < 0.01, ***P < 0.001). B, D Verifications of key protein expression in endoplasmic reticulum signaling including p-eIF2α, GRP78, XBP1, ATF4, DDIT3 and DR5 in response to LINP1 knockout and overexpression by Western blot. E Trypan blue staining and Sytox Green staining were performed to evaluate the cell death induced by LINP1 depletion and overexpression in cSCC cells. Scale bars, 50 μm. F, G Apoptosis assay by Annexin V-APC/7-AAD double staining was performed in cSCC cells after LINP1 knockdown or overexpression. H TUNEL assay was performed to detect apoptosis after LINP1 knockdown or overexpression. Scale bars, 100 μm. I ProCaspase-8, cleaved Caspase-8, proCaspase-7, cleaved Caspase-7, proCaspase-3 and cleaved Caspase-3 were detected by Western blot in cSCC cells after LINP1 depletion or overexpression. *P < 0.05, **P < 0.01, ***P < 0.001)