Fig. 3

The protein abundance of GSDME was negatively regulated by APC/CDC20 but not APC/Cdh1 in prostate cancer cell lines A Scatter plot shows the mRNA expression of GSDME and CDC20 in 12 cell lines derived from the prostate-based CCLE. B The IB analysis of WCL derived from PC-3 D and VCaP cells E infected with the indicated lentiviral shRNA vectors against CDH1. C The IB analysis of WCL derived from PC-3 cells transfected with the constructs of Flag-GSDME, HA-CDC20, and HA-CDH1. D The IB analysis of WCL derived from PC-3 cells transfected with the constructs of Flag-GSDME and HA-CDC20 with or without the addition of MG132. E The IB analysis of WCL derived from PC-3 cells transfected with the constructs of Flag-GSDME, HA-CDC20. F The IB analysis of WCL derived from PC-3 cells transfected with the constructs of Flag-GSDME, wild-type CDC20 (HA-CDC20 WT), and its degrons deleted mutants (HA-CDC20 ΔC-box). G PC-3 cells infected with the indicated lentiviral shRNA constructs against CDC20 for 48 h were selected with 2 µg/ml puromycin for 72 h. The remaining stable cell lines were treated with 100 µg/ml cycloheximide (CHX) and were harvested at the indicated time points. H PC-3 cells were transfected with the indicated constructs. 30 h post-transfection, cells were treated with 100 µg/ml cycloheximide (CHX) and were harvested at the indicated time points. I PC-3 cells infected with the indicated lentiviral shRNA constructs against CDC20 for 48 h were selected with 2 µg/ml puromycin for 72 h. The remaining stable cell lines were treated with or without 10 µg/ml cycloheximide (CHX), and 20 ng/ml TNF-α were harvested at the indicated time points. J PC-3 cells infected with the indicated lentiviral shRNA constructs against CDC20 or GSDME for 48 hours were selected with 2 µg/ml puromycin for 72 h. The remaining stable cell lines were treated with or without 10 µg/ml cycloheximide (CHX), and 20 ng/ml TNF-α were examined with microscopy the 6 h post-treatment. K The quantification of G