Fig. 6: Inhibition of APT2 suppresses ferroptosis.

a Cell viability analysis in A375 cells infected with lentiviruses encoding control, APT1, APT2, ABHD17A, ABHD17B, ABHD17C, and ABHD10 shRNA, followed by 48 h RSL3 treatment. b Cell death analysis in A375 cells infected with lentiviruses encoding control or depalmitoylase shRNAs, treated with RSL3 (4 μM) for 6 h, with or without Fer-1 (2 μM). Phase contrast and fluorescence images (c) and quantitative analysis of dead cells stained with SYTOX Green (d) in A375 cells infected with control or APT2 shRNA and treated with RSL3 (4 μM) for 6 h. Scale bar, 20 μm. e Cell viability analysis in A375 cells pre-treated with ML348, ML349, Palm B, and ABHD957 for 24 h, followed by 48 h RSL3 treatment. f Cell death analysis in A375 cells pre-treated with 2-BP and various depalmitoylase inhibitors for 24 h, followed by RSL3 treatment for 48 h, with or without Fer-1. g Immunoblot confirmation of APT2 and GPX4 expression in A375 cells with APT2 knockdown, infected with HA-APT2 lentivirus. h Cell viability in A375 cells with APT2 knockdown, infected with HA-APT2 lentivirus, followed by 48 h RSL3 treatment. i Immunoblot confirmation of APT2 and GPX4 expression in A375 cells with APT2 knockdown, infected with lentiviruses encoding empty vector, HA-APT2, HA-APT2-C2S, or HA-APT2-S122A. j Cell viability analysis in A375 cells with APT2 knockdown, infected with lentiviruses encoding various HA-APT2 constructs, followed by 48 h RSL3 treatment. k Cell viability analysis in A375 cells infected with control or APT2 shRNA, pre-treated with 2-BP for 24 h, then treated with RSL3 for 48 h. l Cell viability analysis in A375 and HT1080 cells pre-treated with 2-BP and ML349 for 24 h, followed by 48 h RSL3 treatment. Data are presented as mean ± SD. Statistical analysis: two-way ANOVA with Tukey’s test for (b, f, k, l). Sample sizes: (a, b, d–f, h, j–l) n = 3 independent experiments. Image in c, g, i shows representative results from three independent experiments. ns not significant.