Figure 1.

Prtn3 Is Expressed in Hematopoietic Stem/Progenitor Cells and Regulates the Number of Stem and Progenitor Cell Subsets

(A) Prtn3 mRNA expression in sorted BM stem cell-containing populations (LSK cells) in WT and Prtn3^−/− mice as determined by quantitative RT-PCR (n = 3 per group).

(B) Comparison of Prtn3 mRNA expression in sorted LSK cells and neutrophils from WT mice. GAPDH was used as a housekeeping control (n = 3 per group).

(C) PR3 protein expression in sorted BM stem (LSK) and progenitor (LK) cell-containing populations and neutrophils as determined by western blotting. Pan-actin was used as a loading control. Results are representative of three independent experiments.

(D) Intracellular PR3 staining in LSK cells from WT and Prtn3^−/− BM by conventional flow cytometry. Results are representative of five independent experiments.

(E) Intracellular PR3 staining in different cell populations as determined by conventional flow cytometry. Results are representative of five independent experiments.

(F) Representative flow cytometry plots for LSK subsets in WT and Prtn3^−/− mice. Numbers denote the frequency of each population among live singlets.

(G) Quantification of the frequency and number (per one femur and one tibia) of LSK subsets in WT and Prtn3^−/− mice (n = 10 per group).

(H) Representative flow cytometry plots for LK subsets in WT and Prtn3^−/− mice. Numbers denote the frequency of each population among live singlets.

(I) Quantification of the frequency and number of LK subsets in WT and Prtn3^−/− mice (n = 16 per group).

(J) Representative images of histological analysis of BM smears from WT and Prtn3−/− mice. Arrowhead, lymphocyte; long arrow, neutrophil; thick arrow, normoblast; white arrow, monocyte; tailed arrow, myelocyte; right-pointing triangle, myeloblast; wavy arrow, metamyelocyte.

(K) Quantification of cells belonging to different lineages and maturation stages according to histological analysis of BM smears (n = 6 per group).

Scale bar, 10 μm. All values shown are means ± SEM. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, by unpaired, 2-tailed Student's t test. See also Figure S1.