Fig. 5

Knockdown of Atg5 or Atg12 in Tet-On-PADI2 Jurkat T cells inhibits autophagy and induces ER stress and apoptosis. The Tet-On-PADI2 Jurkat T cells with shAtg5 or shAtg12 plasmids were treated with 50 µM Dox for 12 h; “shLuc” was a scrambled RNA knockdown control. A, B The bar graphs illustrate the percentages of shAtg12- and shAtg5-Tet-On-PADI2 cells, respectively, with autophagosomes (orange bars) or apoptotic bodies (green bars) following 12 h treatment with 50 µM Dox. 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. C, D Proteins extracted from shAtg12- and shAtg5-Tet-On-PADI2 cells were detected using antibodies against PADI2, Atg12, Atg5, Atg5–Atg12, LC3B, p62, and β-actin, respectively. E, F Proteins extracted from shAtg12- and shAtg5-Tet-On-PADI2 cells were detected using antibodies against p-PERK, p-eIF2α, CHOP, cleaved-caspase 3, cleaved-PARP and β-actin, respectively. The value indicates the ratio of protein signal to β-actin signal