Fig. 7

CHOP silencing prevents apoptosis and immunogenic cell death induced by lysosome and proteasome inhibitors. MDA-MB-231 cells were transfected with a non-targeting siRNA (scrambled; scr) or with a CHOP-targeting siRNAs pool (siCHOP). Cells were grown in fresh medium (ctrl) or in a medium containing the lysosome inhibitor chloroquine (CQ; 1 μM) or the proteasome inhibitor bortezomib (B; 1 μM), alone or in combination for 24 h. a. Whole cell lysates were probed with the indicated antibodies. The expression of β-tubulin was used as control of equal protein loading. The figure is representative of 1 out of 3 experiments. b. Surface CRT was detected by flow cytometry. The histograms represent the results obtained from 1 out of 3 experiments. Anti-ISO: anti-isotype antibody. c. Tumor cells were stained with PKH2-FITC, DC were stained with an anti-HLA-DR-PE antibody. Tumor cells were co-incubated with DC for 24 h. Double-stained cells were counted by flow cytometry. Data are presented as means±SD (n = 3). *p < 0.001: CQ/B/CQ + B-treated cells vs “scr, ctrl” cells; °p < 0.001: “siCHOP” cells vs corresponding “scr” cells