Extended Data Fig. 8: p150 depletion reprograms H3.3 patterns.

a, Western blotting showing the efficiency of p150 knockdown. Representative images from three independent biological replicate samples are shown. β-actin, loading control. Uncropped blot images are shown as Source Data. b, Genome browser snapshots showing normalized H3.3 enrichment in control and p150-depleted mESCs. Enrichment is shown by log2 ratios between H3.3 and input. c, Genome browser snapshots of the H3.3 ULI-NChIP-seq and H3.3 CUT&RUN data in mESCs. d, Scatter plots showing H3.3 enrichment obtained from H3.3 ULI-NChIP-seq and CUT&RUN in mESCs. The number of 10-kb bins = 252,405. e, Heatmaps showing genome-wide H3.3 enrichment. H3.3 ULI-NChIP-seq and CUT&RUN data obtained by using mESCs were analyzed. The number of 10-kb bins, 252,405. f, Plots showing read enrichment around genic regions. CUT&RUN data using control (normal) IgG was generated for control and p150-depleted mESCs, and read enrichment was analyzed at genic regions. Data from two biological replicates were merged. Note that reads are enriched exactly at the TSSs in p150-depleted mESCs, possibly due to digestion of accessible chromatin by protein A/G-MNase. g, Bar graph indicating relative number of aligned reads. The number of reads from spike-in DNA was used for normalization. h, Plots showing H3.3 enrichment around genic regions. H3.3 CUT&RUN data generated from control and p150-depleted ESCs were analyzed. H3.3 enrichment after spike-in normalization (scaled) is also shown. i, Expression of the indicated transcripts during early mouse development. The published RNA-seq data⁴⁶ was used. j, Quantitative real-time PCR confirmation performed in this study. Gapdh was used for normalization. Log2 fold change of each transcript between control and p150-depleted mESCs are shown. Each dot represents the data from two biological replicates. Error bars indicate mean ± s.d.. Data for graphs are available as source data.

Source data