Fig. 5: PCGF1 contributes to facilitate H3K27me3 deposition and down-regulation of C1 genes in primary HPCs, which safeguards B lineage development. | Nature Communications

Fig. 5: PCGF1 contributes to facilitate H3K27me3 deposition and down-regulation of C1 genes in primary HPCs, which safeguards B lineage development.

From: PCGF1-PRC1 links chromatin repression with DNA replication during hematopoietic cell lineage commitment

Fig. 5

a Box plot of gene expression in each cluster in the control and Pcgf1-KO LMPPs. Data in graphs represent mean for two biologically independent experiments. The center circle indicates a median value and the boxes indicate 25th to 75th percentile. Each dot indicates individual genes. The numbers below the graph are p-values calculated with the Wilcoxon signed rank test. b Gene expression profiling of “Myeloid” related genes in the control and Pcgf1-KO LMPPs. Data in graphs represent mean for two independent biological experiments. Red and blue dots represent C1 and C2 genes, respectively. c Up-regulation of C1 genes in Pcgf1-KO LMPPs shown by qRT-PCR analysis. Data represent mean ± SD of three independent experiments. The numbers on the graph are p-values between the control and Pcgf1-KO calculated with the Welch’s two-sided t test. d A heatmap of ChIP-seq signals for RING1B and H3K27me3 across TSS ( ± 10 kb) of C1 and C2 in control and Pcgf1-KO LSK cells. H3K27me3 ChIP-seq was calibrated by spike-in chromatin. e ChIP-qPCR analyses for local binding of RING1B, H2AK119ub1, SUZ12, and H3K27me3 at selected C1 and C2 genes in the control and Pcgf1-KO LMPPs. Data represent mean ± SD of three independent experiments. The numbers on the graph are p-values between the control and Pcgf1-KO calculated with the Welch’s two-sided t test. f Schematic representation of the experimental procedure. LSK cells sorted from fetal livers of ERT2-Cre;Pcgf1fl/fl or Pcgf1fl/fl mice were infected with either shHmga2-1 or shHmga2-2 retrovirus and treated with 4-OHT for the next 2 days. Eleven days later, the cells were analyzed by FACS. g Flow cytometric profiles for CD19 and Mac1 in respective conditions are shown. Data shown in the graphs are representative of independent biological duplicates. h Partial restoration of total cell number of Pcgf1-KO CD19+ cells in each femur by shHmga2-1. In contrast, the impact of shHmga2-2 was limited. Bar graphs represent mean ± SD of two independent experiments.

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