Fig. 3: LphD and RomA target H3K14 synergistically.
From: Legionella para-effectors target chromatin and promote bacterial replication
Western blot quantification of H3K14 (A), H3K18 or H3K23 acetylation (B). THP-1 cells infected with L. pneumophila (Lp) wild-type (wt) (green) or ∆lphD (white) expressing EGFP. Infected cells were enriched by FACS sorting (EGFP signal). Histones were isolated by acidic extraction and analysed by western blot. Loading control: Histone H1, signal is fold-change normalized to non-infected cells. Data are presented as mean values ± SD of n = 3 independent experiments. For A statistical analyses a two-way ANOVA was performed using Šidák’s multiple comparisons test (*p = 0.0317, **p = 0.0054). C Western blot quantification of H3K14 methylation. THP- 1 cells were infected with Lp wt (green), ∆lphD (white) expressing EGFP. Infected cells were enriched by FACS sorting (EGFP signal). Histones were isolated by acidic extraction and analysed by western blot. Loading control: Histone H1, signal is fold-change normalized to non-infected cells. Data are presented as mean values ± SD of n = 3 independent experiments. For statistical analyses a two-way ANOVA was performed using Šidák’s multiple comparisons test (*p = 0.0136, **p = 0.0010, ****p < 0.0001). D ChIP of H3K14me during Lp infection followed by qPCR targeting selected promoters. THP-1 cells were uninfected (orange) or infected 7 hours with Lp wt (green) or ∆lphD (white) expressing EGFP. Infected cells were enriched by FACS (EGFP signal). Signal was normalized to histone H3. Data are presented as mean values ± SD of n = 3 independent experiments. For statistical analyses a two-way ANOVA was performed using Tukey’s multiple comparisons test with *p as indicated. E ChIP of LphD during Lp infection followed by qPCR targeting selected promoters. THP-1 cells were uninfected (orange) or infected with Lp wt (green) or ∆lphD (white) expressing EGFP. Infected cells were enriched by FACS (EGFP signal). Signal was normalized to histone H3. Data are presented as mean values ± SD of n = 4 independent experiments. All source data in this Figure are provided as source file.
