Figure 2. Reduced proliferation and differentiation of bone marrow cells from Ndrg1 KO mice.

(a) Morphological observations of BM cells at days 2 and 4 after stimulation of M-CSF (left) (original magnification ×40). Comparison of cell proliferation rates between BM cells from WT and Ndrg1 KO mice in the presence of M-CSF (20 ng/mL) (right) (n = 3 per genotype). (b) Dose-response curves of cell proliferation in BM cells in the presence of the indicated concentrations of M-CSF (n = 3 per genotype). (c) Flow cytometric analysis of CD11b+, F4/80+ macrophage population in BM cells at days 2, 4 and 6 in the presence of M-CSF (20 ng/mL) (left). The percentages of CD11b+, F4/80+ cells (macrophages) are shown (right) (n = 3 per genotype). (d) Western blots show time kinetics for the phosphorylation of Erk and Akt in BMDMs at day 2 when cultured in serum free medium for 6 h, and then stimulated with 30 ng/mL M-CSF for indicated time. The right panel shows the quantification of expression levels of p-Erk and p-Akt normalized to the expression levels of GAPDH (n = 3 per genotype). Full-length blot are presented in Supplementary Figure 2. (e) Flow cytometric analysis of CD11c+ dendritic cell population in BM cells at days 3, 5 and 7 in the presence of GM-CSF (25 ng/mL) (left). The percentages of CD11c + cells (dendritic cells) are shown (right) (n = 3 per genotype). (f) Western blots show time kinetics for the phosphorylation of STAT5 in BMDCs at day 2 when cultured in serum free medium for 6 h, and then stimulated with 10 ng/mL GM-CSF for indicated time. The lower panel shows the quantification of expression levels of p-STAT5 normalized to the expression levels of GAPDH (n = 3 per genotype). Full-length blot are presented in Supplementary Figure 3. Each bar is an average ± SD, *P < 0.05; **P < 0.01 versus WT mice (two-tailed Student’s t-test).