Fig. 2

The protein stability of GSDME was negatively regulated by CDC20 through the proteasome pathway. A Box plots the differential expression of pyroptosis-related genes between prostate cancer and normal prostate tissues. Pink color labels prostate cancer (n = 499), and the normal blue controls (n = 52). B, C The immunohistochemical staining (IHC) for protein expression of gasdermin proteins B, which were scored and compared statistically C. D IB analysis of whole cell lysates from PC-3 cells. Cells were harvested at indicated time points after Cycloheximide (CHX) addition with/without MG132 treatment. E Immunoblot (IB) analysis of whole cell lysates from PC-3 cells (left) and VCaP cells (right). Cells were harvested 12 h after the addition of DMSO or MG132 treatment. F The immunoblot (IB) analysis of whole cell lysates (WCL) derived from PC-3 B and VCaP cells C infected with the indicated lentiviral shRNA vectors against CDC20. 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 or without 10 µg/ml cycloheximide (CHX), and 20 ng/ml TNF-α were examined with microscopy the 6 hours post-treatment. H The quantification of G. I Lactate dehydrogenase (LDH) release in the culture supernatants of PC-3 cells infected with the indicated lentiviral shRNA constructs, treated with DMSO, 10 µg/ml cycloheximide (CHX), or 20 ng/ml TNF-α