Fig. 4: STAP-1 regulates apoptosis in CML LSCs.

LSCs (GFP⁺ LSK cells) in BM of primary WT or STAP-1 KO CML mice were analyzed at day 11 after first BMT. a Sorted LSCs were cultured in methylcellulose medium for 7 days, colonies were counted, and cells were serially replated (n = 10 per group). b The cell cycle status of LSCs was analyzed by flow cytometry using Ki67 staining (n = 7 per group). c BrdU was administered to WT or STAP-1 KO CML mice in in vivo as described in Material and Methods to assess the cell cycle status of GFP⁺ Lin⁻ c-Kit⁺ cells with flow cytometry (n = 7 per group). d Cell apoptosis was analyzed by Annexin V and 7AAD in GFP⁺ cells or GFP⁺ LSK cells from primary WT or STAP-1 KO CML mice using flow cytometry. Representative flow cytometric data of the apoptosis are shown in left panel. The numbers in the plot indicate mean percentages ± SD of gated cells. Bar graph represents mean proportion of Annexin V⁺ cells (n = 7 per group). e Cell apoptosis was analyzed by TUNEL staining. Sorted LSCs were fixed and stained with TUNEL (red) and DAPI (blue). Bar graphs represent the proportion of TUNEL-positive cells. Data are representative of one of the two independent experiments with similar results. **p < 0.01 indicates significant difference. NS not significant.