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

Fig. 1. ATF2 and SETDB1-KAP1 dependent induction of ZDHHC20L expression.

Fig. 1

a, b Western blot (WB) of ZDHHC20 isoforms (Short-S, 42 kDa; 20 L, ~49, 62, 69 kDa) and GAPDH (loading control) in a, indicated cancer cell lines and SARS-CoV-2-infected Calu-3 cells (MOI 0.1, 24 h); b primary human airway epithelia from donors of different ages, untreated or SARS-CoV-2-infected (MOI 0.1, 48 h). c Overview of the ZDHHC20 locus (first intron, exon, 5′UTR; hg19). Tracks show in-frame start codons (ATGs), transcript (GENCODE), histone modifications (ENCODE: H3K9me3, repression; H3K4me1, enhancer; H3K27Ac, activation; H3K4me3, promoter), and SETDB1/KAP1, ATF2, SP1, FOXA1 ChIP-seq peaks. d WB as in a, total and phospho-ATF2 in HeLa cells (siCtrl or siATF2), untreated or treated with proaerolysin (10 ng/ml, 1 h at 37 °C, incubated further 8 h). Quantification of 20 L isoforms shown as mean ± SEM, dots represent independent experiments (n = 10); p values by two-way ANOVA, Sidak’s correction. e WB as in a, in U2OS expressing ATF2 (WT, 6D, 6 A), or FOXA1 and SP1. Results mean ± SEM, independent experiments (Control - Ctrl, n = 6; others, n = 3); p values versus Ctrl, two-way ANOVA, Dunnett’s correction. f Cell viability of HeLa cells (ATP assay), normalized to untreated siCtrl (100%), mean ± SD from one of three representative assays (n = 34 replicates); p values vs. siCtrl, two-way ANOVA, Dunnett’s correction. g Schematic: KAP1-KZFPs recruit SETDB1, promoting H3K9me3-mediated repression and active histone mark removal (adapted from ref. 36). h WB as in d, U2OS cells siCtrl, siSETDB1, or siKAP1-depleted (72 h). Results mean ± SEM, independent experiments (n = 6); p values vs. Ctrl, two-way ANOVA, Dunnett’s correction. i WB as in (h) siSETDB1 cells expressing WT or catalytically inactive SETDB1 mutant (Mut). j WB as in a, in HAP-1 wild-type or KAP1-knockout cells. k mRNA quantification across ZDHHC20 transcripts (coding regions: C1, C2; 5’UTR positions: 1–5) in HAP-1 wild-type or KAP1-knockout cells, mean ± SEM, biologically independent experiments, p values vs. normalized WT, two-way ANOVA, Dunnett’s correction. Source data and entire blots provided as a Source Data file and supplementary information.