Extended Data Figure 1: EZH1 knockdown activates lymphoid potential from pluripotent stem cells.

a, List of all candidate epigenetic modifiers in loss-of-function shRNA screen. b, Representative flow plots of CD4⁺CD8⁺ T cell potential across top six candidates from four independent hairpins in two independent experiments (n = 8). See Fig. 1. c, CD34⁺ cells were isolated after 9 days of embryoid body (EB) differentiation (top left), transduced with shLUC or shEZH1 and cultured under conditions that promote endothelial-to-haematopoietic transition⁴⁴. After 6 days, rounded haematopoietic cells (top right) were collected and co-cultured on OP9-DL1 stroma. Bottom, flow cytometric analysis of T cell potential in shLUC and shEZH1 cells without 5F is shown for two independent iPS lines (34-iPS and MSC-iPS1) in one experiment (n = 2 biological replicates). PSC-HE, pluripotent stem-cell-derived haemogenic endothelium. d, Expansion and differentiation potential of 5F plus shEZH1 cells after long-term in vitro culture. 5F plus shEZH1 cells were maintained in cultures containing doxycycline for 14 days respecification (approximately 100-fold expansion), plus an additional 6 weeks (approximately 1,000-fold expansion) and then plated into OP9-DL1 stromal cells for T cell differentiation. Representative flow cytometric analyses of T cell potential of 5F plus shLUC and 5F plus shEZH1 cells after 13 weeks of expansion and differentiation (n = 2 biological replicates). e, Flow cytometric analysis (left) and quantification (right) of the proportion of CD34⁺ and CD34⁻ haematopoietic progenitors in doxycycline-containing suspension culture at day 25 (n = 2 biological replicates).

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