Fig. 1
H2Aub directly contributes to the deposition of H3K27me3. a Lysine residues in Arabidopsis and Marchantia H2A. Lysine 120 (K120) and K121 of H2A in Arabidopsis and potential ubiquitination sites (K115, K116, and K119) in H2A of Marchantia are highlighted. b Western blot of bulk H2Aub and H2A in wild type (WT), Mph2a;H2AK115R/K116R #1, Mph2a;H2AK115R/K116R #2, Mph2a;H2AK119R #1, and Mph2a;H2AK119R #2 mutants. c–e Phenotypes of 28-day-old WT, Mph2a;H2AK115R/K116R #1, and Mph2a;H2AK119R #1 mutants. Blue arrows point to downward curled edges of the thallus. Scale bars, 1 cm. f–h Gemma cup phenotypes of WT, Mph2a;H2AK115R/K116R #1, and Mph2a;H2AK119R #1 mutants. Blue arrowheads point at visible rhizoids produced by non-dormant gemmae in the mutants. Scale bars, 0.05 cm. i Boxplots showing H2Aub and H3K27me3 levels (RPKM, reads per kilobase per million mapped reads) of genes losing H2Aub in Mph2a;H2AK115R/K116R in WT, Mph2a;H2AK115R/K116R, and Mph2a;H2AK119R mutants. j Boxplots showing H2Aub and H3K27me3 levels (RPKM) of genes losing H2Aub in Mph2a;H2AK119R in WT, Mph2a;H2AK119R, and Mph2a;H2AK115R/K116R mutants. k Boxplot showing H3K27me3 levels (RPKM) of genes that do not show significant changes of H2Aub in h2a_ub mutants in WT, Mph2a;H2AK115R/K116R, and Mph2a;H2AK119R mutants. H2Aub and H3K27me3 levels in i-k were calculated as the average RPKM from 1 kb upstream of the transcriptional start to the transcriptional end of genes. l Venn diagram showing overlap of H2Aub-marked genes and H3K27me3-marked genes in wild type (WT). Significance was tested using a hypergeometric test. m Boxplot showing H3K27me3 levels of H2Aub/H3K27me3 genes in WT, Mph2a;H2AK115R/K116R, and Mph2a;H2AK119R mutants. H3K27me3 levels were calculated as the average RPKM from 1 kb upstream of the transcriptional start to the transcriptional end of genes. n Venn diagram showing overlap of H2Aub-marked transposable elements (TEs) and H3K27me3-marked TEs in WT. o Boxplot showing H3K27me3 levels of H2Aub/H3K27me3 TEs in WT, Mph2a;H2AK115R/K116R, and Mph2a;H2AK119R mutants. H3K27me3 levels were calculated as the average RPKM from the start to the end of TEs. Boxes show medians and the interquartile range, and error bars show the full range excluding outliers. **p < 0.01 (Wilcoxon test)