Extended Data Fig. 7: GATA3 induction upon pharmacological and genetic targeting of PRC2.
a Immunofluorescence microscopy (on Olympus IX81, 20x magnification) showing H3K27me3 and GATA3 staining of naïve HS975 hESC cultured in 5iLAF medium with or without EZH2i (EPZ-6438) treatment for 7 days. Scale bars are 50 µm. Tukey boxplot generated from corresponding CellProfiler image analysis shows per-nucleus H3K27me3 and GATA3 intensities over n = 195 nuclei (69 untreated, 127 EZH2i) derived from 1 experiment. EZH2i treatment resulted in 6.3% GATA3 + nuclei for this cell type. The threshold for GATA3 + cells (dashed line) is defined as 1.5x mean of untreated cells. Box plot boxes show the 25th and 75th percentile with the median, and whiskers indicate 1.5x the interquartile range. P values estimated by two-sided unpaired t test (p = 0.047; p < 2.2e-16, respectively). b Boxplots show per-nucleus EED (top panel) and GATA3 (bottom panel) intensities summarized by the imaged colonies for CRISPR/Cas9 based acute targeting of EED. corresponding to Fig. 4f. Box plot boxes show the 25th and 75th percentile with the median, and whiskers indicate 1.5x the interquartile range. c Immunofluorescence microscopy showing H3K27me3 and GATA3 staining of naïve H9 hESC cultured in t2iLGö (NaiveCult) or PXGL medium with or without EZH2i (EPZ-6438) treatment for 7 days. Scale bar is 20 µm. Tukey boxplot generated from corresponding CellProfiler image analysis shows per-nucleus H3K27me3 and GATA3 intensities over 3045 nuclei in total (500 t2iLGö, 846 t2iLGö +EZH2i, 1037 PXGL, 662 PXGL + EZH2i) derived from 1 experiment. The threshold for GATA3 + cells (dashed line) is defined as 1.5x mean of untreated cells. Box plot boxes show the 25th and 75th percentile with the median, and whiskers indicate 1.5x the interquartile range.
