Fig. 3

Overexpressing PADI2 increases citrullinated proteins and autophagy-related proteins expression in activated Jurkat T cells. A Jurkat Tet-On-Vector and Tet-On-PADI2 cells were treated with 50 μM Dox for 12 h. Anti-citrulline antibodies were used to immunoblot lysates from cells. B Proteins extracted from Tet-On-Vector and Tet-On-PADI2 cells were detected using antibodies against PADI2, p62, LC3B, and β-actin as indicated time after 50 μM Dox exposure. C Proteins extracted from Tet-On-Vector and Tet-On-PADI2 cells were detected using antibodies against PADI2, Beclin-1, Atg7, Atg5, Atg12, Atg5–Atg12, and β-actin at the indicated time points after 50 μM Dox exposure. D Representative AO staining images of Tet-On-Vector and Tet-On-PADI2 cells with 50 μM Dox treatment for 12 h, scale bar: 20 µm. The cells exhibited autophagosomes as indicated by arrowheads. E The bar graphs show the percentages of cells with autophagosomes with 50 μM Dox treatment for 12 h. The data are presented as the mean ± SEM of three separate experiments (***p < 0.001). The p value in was calculated by two-way ANOVA. F Following 30 µM CQ treatment, proteins extracted from Tet-On-Vector and Tet-On-PADI2 cells were detected using antibodies against LC3B, p62, cleaved caspase-3, cleaved-PARP and β-actin at the indicated time points after 50 μM Dox exposure for 24 h. G Proteins extracted from Tet-On-Vector and Tet-On-PADI2 cells were detected using antibodies against LAMP1, LAMP2 and β-actin at the indicated time points after 50 μM Dox exposure for 24 h. The value indicates the ratio of protein signal to β-actin signal