Fig. 3

Fas is not required for IFNγ/TNFα-induced apoptosis in MSCs. a Percentage of Fas^−/− BM-MSC cell survival after treatment with various concentrations of different cytokines. b Fas^−/− BM-MSCs were treated with the indicated cytokines for 48 h, apoptotic cells were analyzed using flow cytometry. The percentage of Annexin V-positive cells relative to untreated controls is indicated in a bar chart. c Fas^−/− BM-MSCs were treated with the indicated cytokines for 24 h; pro-apoptotic and anti-apoptotic transcripts were quantified by real-time PCR. d Fas^−/− BM-MSCs were treated with IFNγ/TNFα (10 ng/ml each) in the absence or presence (1 h pre-incubation) of L-NMMA (1 mM) for 48 h. Apoptotic cells were analyzed using flow cytometry. The percentage of Annexin V-positive cells relative to untreated controls is indicated in a bar chart. e Fas^−/− BM-MSCs were treated as described in d. Pro-apoptotic and anti-apoptotic transcripts were quantified by real-time PCR. f Fas^−/− BM-MSCs were treated with SNAP (0.5 mM) or IFNγ/TNFα (10 ng/ml each) for 48 h. Apoptotic cells were analyzed using flow cytometry. g Fas^−/− BM-MSCs were treated with SNAP (0.5 mM) for 24 h; pro-apoptotic and anti-apoptotic transcripts were quantified by real-time PCR. *P < 0.05, **P < 0.01, ***P < 0.001