Fig. 8.

PANX1 mediates chemotherapy drugs-induced lytic cell death independent of gasdermins and MLKL. A: Representative images of WT, ABCDEM-6KO, PANX1-deficient, and PANX1-deficient in 6KO HeLa-pTet-nSREBP2 cell treated with or without 10 μM doxorubicin or 30 μM cisplatin for 24 h. Cells were preloaded with PI for 15 min and imaged at 63× magnification. Scale bar, 25 μm. B: representative images of ABCDEM-6KO and PANX1-deficient in 6KO HeLa-pTet-nSREBP2 cell treated with or without cisplatin for 24 h. Scale bar, 25 μm. C: LDH release test. Data are presented as means ± SD. Statistical analyses, unpaired two-tailed Student’s t test. D and E: the ABCDEM-6KO and PANX1-deficient HeLa-pTet-nSREBP2 cells were transfected with a plasmid expressing wild-type or mutated PANX1 (D379A). All cells were treated with doxorubicin (D) or cisplatin (E) for 24 h. Data are presented as means ± SD. Statistical analyses, unpaired two-tailed Student’s t test. F: cell viability test. HeLa cells were plated in 10% FBS medium or sterol depletion medium (5% lipoprotein-deficient serum, 1 μM lovastatin, 10 μM mevalonate) for 16 h, then treated with indicated concentrations of doxorubicin (starting from 15 μM, with 3-fold serial dilutions) for 24 h. Nonlinear regression was performed in Prism and EC₅₀ was calculated. G: LDH release test. HeLa-pTet-nSREBP2 cells were plated with 0/0.03 μg/ml doxycycline for 8 h, then treated with indicated doxorubicin for 24 h. H, Proposed model of nSREBP2 induced lytic cell death. ∗P < 0.05, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. ns, not significant.