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. 2020 Jul 9;15(2):326–339. doi: 10.1016/j.stemcr.2020.06.008

Figure 6.

Figure 6

Variations in GATA2 Protein Levels Correlate with HSPC Proliferation and Lineage Potential

(A) Quantification of liquid colonies from single preGMs or GMPs by quantitative imaging of cell morphology, nuclear shape, and expression of GATA2VENUS, CD115 (monocytes), LY6G (neutrophils), and FCεR1 (mast cells). Examples shown for monocyte, neutrophil, mast cell, bipotent monocyte-neutrophil, bipotent mast cell-neutrophil, and tripotent monocyte-neutrophil-mast cell colonies. Immature colonies do not express any surface markers. Scale bar, 50 μm.

(B) Four different preGM and GMP fractions based on GATA2 levels were single-cell sorted into 384-well plates and cultured for 8 days in pan-myeloid media (IMDM + FCS + BIT + SCF + GM-CSF + IL-3 + IL-9).

(C) GATA2 -low and -mid preGMs exhibit higher proliferation and colony-forming potential than GATA2 -negative and -high cells. Data represented by box and whisker plots with median. Dots indicate individual measurement per cell (n = 3 independent mice).

(D) GATA2 protein expression correlates with different preGM and GMP lineage potential. Mean percentage of different colony types identified in (A) (n = 3 independent mice). Error bars = SD. Number of colonies, preGMs: GATA2-neg, 155; GATA2-low, 225; GATA2-mid, 157; GATA2-high, 152. GMPs: GATA2-neg, 173; GATA2-low, 153; GATA2-mid, 161; GATA2-high, 91. Two-sided Wilcoxon rank-sum test; ∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05.