Fig. 5

Proliferation of nuclear OCT-4⁺ VSELs. a Dual immunofluorescence on cryosections of D4 femur detected the presence of nuclear OCT-4⁺ cells co-expressing proliferation marker PCNA. Nuclear OCT-4/PCNA co-expressing VSELs were observed close to the endosteal region. The nuclei were counterstained with DAPI. Scale bar = 100 μm. Inset: magnified image of cluster of OCT-4/PCNA coexpressing cells. Scale bar = 10 μm. b, c Results of the CFU assay: small and large hematopoietic colonies of type CFU-GM or CFU-GEMM were observed on culture of cells that survived 5-FU on D4 in Methocult media. d GFP⁺ cell clusters observed after 10 days of culture of 5-FU-treated GFP cells in Methocult media. e Typical cobblestone formation representative of hematopoietic colonies seen in 5-FU-treated cell culture. f Representative image of colonies observed in CFU assay at 20× magnification, colony formed is of the type CFU-GEMM. OCT-4 octamer binding transforming factor-4, PCNA proliferating cell nuclear antigen, DAPI 4′,6-diamidino-2-phenylindole, CFU-GM colony-forming unit granulocytes/macrophage, CFU-GEMM colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte