HTA9 | Probable histone H2A variant 3

A modified form of H2A.Z is required to regulate flowering time. a Western blot (WB) analysis of HTA9 levels in histone enriched extracts of wild-type (WT) and hta9hta11 mutants at 7 days after germination (DAG; upper panel). The same blot was probed with anti-H4 antibody for loading control (bottom panel). Arrows indicate the bands recognized by the antibody. b Schematic representation of the C-terminal region of FLAG-tagged HTA9 native construct (FLAG-HTA9_N) or mutated in which K129 or both K129 and K132 were replaced by R (FLAG-HTA9_RK and FLAG-HTA9_RR, respectively). c WB analysis of HTA9 and FLAG-HTA9 levels in WT, hta9hta11, WT/FLAG-HTA9_N, WT/FLAG-HTA9_RK, and WT/FLAG-HTA9_RR (upper panel). Arrows indicate the bands recognized by the antibody. The same blot was probed with anti-H4 antibody for loading control (bottom panel). d Picture showing WT, hta9hta11, hta9hta11/FLAG-HTA9_N, hta9hta11/FLAG-HTA9_RK, and hta9hta11/FLAG-HTA9_RR plants at 24 DAG growing under long day conditions. Arrows indicate the presence of elongated shoot. Bar indicates 1?cm. e Box plots indicating the number of rosette leaves at bolting in different genotypes. “n” indicates the number of plants. In each case, the median (segment inside rectangle), upper and lower quartiles (boxes), and minimum and maximum values (whiskers) are indicated. p Value of significant differences as determined by Student’s t test are indicated. f Relative FT expression levels in the different genotypes at 12 DAG. ACTIN2 (ACT2) was used as an internal control. Error bars indicate standard deviation of n?=?4 biological replicates. Data points are indicated in the bar charts. g Relative FLC expression levels in different genotypes at 12 DAG. ACT2 was used as an internal control. Error bars indicate standard deviation of n?=?4 biological replicates. Data points are indicated in the bar charts. Significant differences as determined by Student’s t test are indicated (**P?

A modified form of H2A.Z is required to regulate flowering time. a Western blot (WB) analysis of HTA9 levels in histone enriched extracts of wild-type (WT) and hta9hta11 mutants at 7 days after germination (DAG; upper panel). The same blot was probed with anti-H4 antibody for loading control (bottom panel). Arrows indicate the bands recognized by the antibody. b Schematic representation of the C-terminal region of FLAG-tagged HTA9 native construct (FLAG-HTA9_N) or mutated in which K129 or both K129 and K132 were replaced by R (FLAG-HTA9_RK and FLAG-HTA9_RR, respectively). c WB analysis of HTA9 and FLAG-HTA9 levels in WT, hta9hta11, WT/FLAG-HTA9_N, WT/FLAG-HTA9_RK, and WT/FLAG-HTA9_RR (upper panel). Arrows indicate the bands recognized by the antibody. The same blot was probed with anti-H4 antibody for loading control (bottom panel). d Picture showing WT, hta9hta11, hta9hta11/FLAG-HTA9_N, hta9hta11/FLAG-HTA9_RK, and hta9hta11/FLAG-HTA9_RR plants at 24 DAG growing under long day conditions. Arrows indicate the presence of elongated shoot. Bar indicates 1?cm. e Box plots indicating the number of rosette leaves at bolting in different genotypes. “n” indicates the number of plants. In each case, the median (segment inside rectangle), upper and lower quartiles (boxes), and minimum and maximum values (whiskers) are indicated. p Value of significant differences as determined by Student’s t test are indicated. f Relative FT expression levels in the different genotypes at 12 DAG. ACTIN2 (ACT2) was used as an internal control. Error bars indicate standard deviation of n?=?4 biological replicates. Data points are indicated in the bar charts. g Relative FLC expression levels in different genotypes at 12 DAG. ACT2 was used as an internal control. Error bars indicate standard deviation of n?=?4 biological replicates. Data points are indicated in the bar charts. Significant differences as determined by Student’s t test are indicated (**P?

The AtBMI1 proteins mediate H2A.Z monoubiquitination. a Western blot (WB) analysis of HTA9 levels in histone-enriched extracts of wild-type (WT) and atbmi1a/b/c mutant at 7 days after germination (DAG). The same blot was probed with anti-H4 antibody for loading control. b Quantification of HTA9 and HTA9ub levels in WT and atbmi1a/b/c mutant relative to H4; Error bars indicate standard deviation of n?=?3 independent experiments. Data points are indicated in the bar charts. c WB analysis of WT and atbmi1a/b/c mutant histone-enriched extracts at 7 DAG probed with anti-ubiquitin antibody. Bands corresponding to H2Bub, H2AK121ub, and HTA9ub are indicated. A blot probed with anti-H4 antibody was used as a loading control. d WB analysis of HTA9 and FLAG-HTA9 levels in WT, atbmi1a/b/c, WT/FLAG-HTA9_N, and atbmi1a/b/c/FLAG-HTA9_N. Arrows indicate the bands recognized by the antibody. The same blot was probed with anti-H4 antibody for loading control. Source data of Fig. 2a–d are provided as a Source Data file

The AtBMI1 proteins mediate H2A.Z monoubiquitination. a Western blot (WB) analysis of HTA9 levels in histone-enriched extracts of wild-type (WT) and atbmi1a/b/c mutant at 7 days after germination (DAG). The same blot was probed with anti-H4 antibody for loading control. b Quantification of HTA9 and HTA9ub levels in WT and atbmi1a/b/c mutant relative to H4; Error bars indicate standard deviation of n?=?3 independent experiments. Data points are indicated in the bar charts. c WB analysis of WT and atbmi1a/b/c mutant histone-enriched extracts at 7 DAG probed with anti-ubiquitin antibody. Bands corresponding to H2Bub, H2AK121ub, and HTA9ub are indicated. A blot probed with anti-H4 antibody was used as a loading control. d WB analysis of HTA9 and FLAG-HTA9 levels in WT, atbmi1a/b/c, WT/FLAG-HTA9_N, and atbmi1a/b/c/FLAG-HTA9_N. Arrows indicate the bands recognized by the antibody. The same blot was probed with anti-H4 antibody for loading control. Source data of Fig. 2a–d are provided as a Source Data file

Overlap between upregulated genes in hta9hta11 and atbmi1a/b. a Venn diagram showing the number of overlapping genes when comparing H2A.Z-enriched genes in wild-type (WT) seedlings (H2A.Z_WT), upregulated genes in hta9hta11, and upregulated genes in atbmi1a/b weak mutant. b Metagene plots showing H2A.Z levels over different subsets of genes grouped according to their expression levels (indicated in FPKM (fragments per kilobase of exon and million mapped reads) and different colors). TSS transcription start site, TES transcription stop site. c Metagene plots showing H2A.Z enrichment in WT at genes upregulated in hta9hta11 and atbmi1a/b weak mutants and at genes downregulated in hta9hta11. d Venn diagram showing the number of overlapping genes when comparing H2A.Z-enriched genes in WT (H2A.Z_WT), H2AK121ub marked genes in WT (H2AK121ub_WT), and upregulated genes in hta9hta11. e WB blot showing H2AK121ub and HTA9ub levels in WT, hta9hta11, and atbmi1a/b/c (right and left panels, respectively). The blot was probed with an anti-AtH2AK121ub antibody, then stripped and re-probed with anti-HTA9 antibody. Arrows indicate the bands recognized by the antibody. The same blot was probed with anti-H4 antibody for loading control (bottom panel). Source data of Fig. 3e are provided as a Source Data file