G-CSF–independent and -dependent CH HSPC defects. (A) Messenger RNA quantification from FACS-isolated HSC (LSK CD150+ CD48−) or MPP (LSK CD150– CD48–). N = 3 to 6 per condition from 3 experiments. (B) CFU from 2 × 105 total BM cells or 0.1 mL of peripheral blood. N = 3 to 4 per condition from 3 experiments. ∗P < .05, ∗∗P < .01; 1-way ANOVA followed by Tukey multiple comparisons test. (C) Top: EdU incorporation to quantify cell entry into S phase within HSCs and MPPs. Bottom: representative flow cytometric plots. N = 4 to 7 per condition, 5 experiments. (C) Top: annexin V binding to quantify HSC and MPP apoptosis. Bottom: representative flow cytometric plots. N = 4 to 7 per condition, 5 experiments. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001; 2-tailed unpaired Student t test with Benjamini-Hochberg correction. DAPI, 4′,6-diamidino-2-phenylindole; FACS, fluorescence-activated cell sorter.
