Fig. 10

PADI2 silencing has the opposite effect on PADI2-induced ER stress, autophagy, and Th17-like T cell activation in Tet-On-PADI2 Jurkat T cells. The Tet-On-PADI2 Jurkat T cells with shLuc or shPADI2 plasmids were treated with 50 µM Dox for 24 h or 48 h; “shLuc” was a scrambled RNA knockdown control. A The viability of shLuc- and shPADI2-Tet-On-PADI2 cells with 50 µM Dox is presented as the mean ± SEM of three independent experiments (**p < 0.01, and ***p < 0.001). The p value in was determined by one-way ANOVA. B, C The bar graphs depict the percentages of shLuc- and shPADI2-Tet-On-PADI2 cells with autophagosomes (orange bars) or apoptotic bodies (green bars) after 24 or 48 h of 50 µM Dox treatment, respectively. The data are presented as the mean ± SEM of three separate experiments (***p < 0.001). The p value in was calculated by one-way ANOVA. D Proteins extracted from shLuc- and shPADI2-Tet-On-PADI2 cells were detected using antibodies against BECN1, LC3B, p62, and β-actin. E Proteins extracted from shLuc- and shPADI2-Tet-On-PADI2 cells were detected using antibodies against PADI2, p-PERK, p-eIF2α, cleaved-PARP, and β-actin. F RT-PCR was used to determine the levels of IL-6, IL-17A, IL-21, IL-22, and β-actin mRNA expression. The value indicates the ratio of protein signal to β-actin signal