Fig. 2: Domino is the main histone chaperone for H2A.Z on TSS.
From: Histone variant H2A.Z regulates zygotic genome activation
a Experimental setup for H2A.Z IP-MS. b Volcano plot showing the -log10 p-value (y axis) compared to the enrichment over control (log2 LFQ) of the anti-Flag IP followed by MS. A fitted linear model with padj for multiple-hypothesis (Benjamini–Yekutieli, two-sided) was used to obtain significantly enriched or depleted proteins, colored in cyan (-log10 padj value >1.3). The red dotted line frames significantly enriched proteins (log2 LFQ > 0). Domino isoforms are highlighted in magenta. Dark blue denotes significantly enriched members of the Tip60 complex. Yellow is Arp6, a DominoB34 interactor. n = 3 biological replicates per condition. c Representative immunofluorescence of DominoB-GFP in pre-ZGA (left, NC10) and ZGA (right, NC14) embryos. Insets show a magnification of the GFP signal. Scale bar = 40 μm. See Supplementary Fig. 2a for DomA staining. d Phenotypic characterization of Ctrl and DomKD embryos. Percentage of embryos (left) reaching the ZGA stage or (right) completing embryogenesis (Hatching) (p-value < 0.0001, two-tailed Mann–Whitney test). n = 453 embryos for Ctrl and n = 465 embryos for DomKD embryos. Data are presented as mean values ± SD. Source data are provided as a Source Data file. e Violin plot of the mean rlog transformed counts of H2A.Z quantitative ChIP-seq on active promoter regions in Ctrl and DomKD embryos at ZGA. Box plot inside depict the median and the interquartile range (IQR) from the 1st to the 3rd quartile. Whiskers indicate the upper (Q3 + 1.5*IQR) and lower edge (Q1 − 1.5*IQR). Welch two sample t-test, two-sided, p-value < 2.2e−16. n = 2 biologically independent experiments per genotype. f Heatmap of H2A.Z quantitative ChIP-seq signal (left 2 columns) and Zelda Cut&Tag signal (right 2 columns) on active promoters, sorted according to H2A.Z signal on Ctrl embryos at ZGA. Averaged library size corrected signal tracks from two replicates were used for Zelda. g Screenshots of genome browser tracks (Integrative Genomics Viewer, IGV)114 showing H2A.Z quantitative ChIP-seq signal in Ctrl and DomKD embryos at ZGA, Rpb3 signal in Ctrl embryos represents RNA Pol II coverage on the same regions. (Top) Chr3L:17,010,000–17,060,000. (Middle) Chr3R: 25,243,000–25,265,000. (Bottom) ChrX: 3,680,000–3,715,000.
