Fig. 5

NO-mediated CD34 expression is independent of the activation of the cGMP signaling pathway, but depends on S-nitrosylation of proteins. Sort-purified LSK-CD34⁻ cells were exposed to 200 μM sodium nitroprusside (SNP) or 100 μM 8-Cl-cGMP, a membrane-permeable cGMP analogue, for 12 h. The cells were analyzed by flow cytometry. a The frequency of LSK-CD34⁺ cells generated in the culture is graphically illustrated. b, c The graph and the flow panel show mean fluorescence intensity (MFI) of CD34 fluorescence. Sort-purified CD34⁻ cells were exposed to 200 μM SNP in the presence or absence of 100 μM Rp-cGMP, a specific inhibitor of cGMP-dependent protein kinase, for 12 h. The cells were subjected to flow cytometry analysis. d The frequency of LSK-CD34⁺ cells generated in the cultures is illustrated. e, f The graph and the flow panel show the MFI of CD34 fluorescence. Sort-purified CD34⁻ cells were exposed to 200 μM SNP in the presence or absence of 200 μM DTT for 12 h. g The frequency of LSK-CD34⁺ cells generated in the cultures is depicted. h, i The graph and the flow panel show the MFI of CD34 fluorescence. The data are represented as mean ± SEM. N = 3. ***p ≤ 0.001. Also see Additional file 5 (Figure S3). NS not significant