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. 2025 Dec 11;17:696. doi: 10.1038/s41467-025-67304-4

Fig. 3. p38 and JNK phosphorylation of ZNF354A.

Fig. 3

a Immunoprecipitation (IP) of ZNF354A-HA from U2OS cells (siZNF354A, 72 h) expressing WT or phospho-mutants of ZNF354A, untreated or treated with proaerolysin (10 ng/ml, 1 h at 37 °C, washed, incubated additional 8 h). Blots show phospho-Ser/Thr proteins and ZNF354A in IP fractions. b Phospho-Tag Western blot of total cell extracts (TCE) from U2OS cells treated as in (a). c Western blot of ZDHHC20 (Short-S, 20 L isoforms), phospho-KAP1 (S473), phospho-ATF2 (T69), total KAP1, ATF2, and GAPDH (loading control) in TCE from U2OS cells treated with proaerolysin as in (a), plus kinase inhibitors. d Western blot as in (c) from TCE and IP-ZNF354A of U2OS cells (siATF2, 72 h) expressing ATF2-6D phosphomimetic mutant (24 h), treated with kinase inhibitors. e Quantification of phospho-KAP1 and phospho-Ser/Thr-ZNF354A bands from blots in (d). Mean ± SEM; dots represent independent experiments (n = 4). Statistical analysis by two-way ANOVA, Dunnett’s test; P values compare treated conditions to untreated control (Ctrl). Source data and entire blots provided as a Source Data file and supplementary information.