Figure 4. TNF increases the number of MSCs with high Notch activation.
(A–C) Hes1-GFP mice were injected with TNF (0.5 μg/injection, ip, twice a day for 5 days) or PBS. The frequency of bone marrow CD45−/GFP+ MSCs (A), the number of CFU-ALP+ colonies (B), and their expression levels of Hes1 were determined by flow cytometry, CFU-ALP colony assays or qPCR. Values are mean ± SD of 4 mice. *p <0.05 vs. PBS-injected mice. (D–G) 6-month-old Hes1-GFP/TNF-Tg mice and Hes1-GFP littermate controls were used. The frequency of bone marrow CD45−/GFP+ MSCs (D), and their expression of Hes1 (E) were determined. Values are mean ± SD of 5 mice. (F) CD45−/GFP+ cells were sorted by FACS and cultured in CFU-ALP+ colony assays. CFU-ALP+ colony #/well were counted. Values are mean ± SD of 3 wells. (G) Bone marrow cells were cultured in CFU-ALP colony assays. The # and % of GFP+ cells/colony were assessed as in Figure 2. Values are mean ± SD of 3 wells. *p <0.05 vs. cells from Hes1-GFP mice.