EZH2 inhibition alters proplatelet formation. (A) Proplatelet formation analysis in cells treated with GSK343 or shEZH2 on day 13 of culture. Means of 3 independent experiments are shown. Data were compared using a t test with Mann-Whitney correction. PPT, proplatelet. All data are shown as mean ± SEM. *P < .05. (B-C) Transcriptome analysis performed on CD41⁺CD42⁺ MKs cultured either in the presence GSK inhibitors (GSK126 or GSK343) or in their absence (CTL) and sorted on day 13 of culture. (B) Volcano plot showing differentially expressed genes in the CD41⁺CD42⁺ cell population in CTL vs treated cells (P < .001). (C) GO analysis of cellular component on the 59 genes common to both inhibitors. Corrected P values were calculated using a modified Fisher’s exact test (P < .001, fold change >2). (D) ChIP-seq profiles of the 3 histone marks (H3K4me3, H3K27me3, and H3K27ac) across FSCN1 on day 12 of MK culture (blue, control; red, GSK343 treated) are shown in the upper panel and ChIP-EZH2 qPCR in the lower panel. One representative ChIP-qPCR experiment of 3 experiments is shown in duplicate on the left, and the statistical analysis of 3 independent ChIP experiments is shown on the right. Data were compared using a Student t test with Mann-Whitney correction. All data represent means ± SEM. *P < .05. (E) Immunofluorescence staining of F-actin (green) and nucleus (blue) in control (CTL) and GSK126-treated MKs on day 13 of culture after adhesion on poly-L-lysine. Scale bars, 30 μm.