Extended Data Fig. 3: ThPOK expression and effect of ThPOK deficiency on gene expression.
From: ThPOK is a critical multifaceted regulator of myeloid lineage development
a) GFP expression by indicated subsets from ThPOK-GFP reporter mice. GFP expression level is color-coded according to heat-scale below each plot. b) Gating strategy to reveal granulocyte-monocyte lineage development (according to REF. 17). Red arrows indicate sequential gates. c) Unsupervised clustering (viSNE) of cells in the modified-GMP gate (upper left). Other panels show expression of indicated surface marker superimposed on the same clusters. d) GFP protein (top), GFP mRNA (center), and endogenous ThPOK mRNA levels, in indicated progenitor subsets sorted according to gating strategy in (b), as well as indicated control T cell populations from transgenic ThPOK-GFP reporter mice. e) Competitive reconstitution assay with WT and ThPOK − /− BM cells (same experiment as depicted in Fig. 1j) (n = 6, per genotype). Relative contribution to indicated BM populations was assessed at 6 months post-transplant. Data are normalized for only donor cells, so that the sum of WT and KO donor cells equals 100%. Two sided unpaired T test with Welch’s Correction was performed. Data are displayed as mean ± SEM (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). f) Cell cycle analyses of indicated gated subsets in WT and ThPOK -/- BM 20 h after a single in vivo BrdU pulse. Representative FACS plots of BM subsets from PBS-injected WT controls (BrdU-), or WT and ThPOK-/- mice that received BrdU, as indicated (right panels). Cell cycle stages were defined by flow cytometry based on BrDU intensity and DNA content (7-AAD staining), as indicated. Bar graphs at left summarize the results for indicated progenitor populations, expressed as means ± SEM (n = 3 independent animals per genotype) and are representative of two experiments. Two sided unpaired t-test with Welch’s Correction was performed (*p < 0.05; **p < 0.01; *** p < 0.001).
