Fig. 7: Ninj1 suppresses Caspase-9-dependent intrinsic apoptosis in preOCs, and incremental Ninj1 expression enhances the survival of preOCs.

a BMMs were cultured with M-CSF and RANKL for the indicated days. Cell lysates were subjected to immunoblot analysis. b RAW264.7 cells transfected with siScr or siNinj1 were cultured with RANKL. Cells were harvested on the indicated days and subjected to immunoblot assays. c GFP⁺ or DsRed⁺ RAW264.7 cells were transfected with scrambled siRNA or Ninj1 siRNA as indicated. An equal number of each fluorescence protein-labeled cell population was cocultured with RANKL. The ratio of GFP⁺ to DsRed⁺ cells was determined using FACS, and the data are shown as the mean ± SD (n = 3, with triplicates in each experiment); **P < 0.01, and ***P < 0.001. d–f RAW264.7 cells were stably transduced with Ninj1-FLAG or empty vector using a retrovirus and cultured with RANKL. Cells were harvested and stained with APC-Annexin V and 7AAD on the indicated days followed by FACS analysis. d Representative plots showing the results of FACS analysis on days 2 and 3. e FACS-determined statistical stacked bars (n = 4). f FACS-determined percent frequency of Annexin V⁺ cells. The data are shown as the mean ± SD (n = 4); **P < 0.01