Figure 3
Characterization of BCC subsets. ALDH1 activity was assessed in positive control with K562 cells (A) and experimentally, with heterogeneous MDA-MB-231 cells (B). The presence of Oct4-dsRED-SB in MBA-MB-231 cells was determined by Bix treatment in flow cytometry (C). Different subsets of Oct4-dsRED-SB-transfected MDA-MB-231 cells (D) were assessed for ALDH1 (E). Similar sorting of three MDA-MB-231 subsets were analyzed for telomere length using the TeloTAGG kit (F). MDA-MB-231, stably transfected with Oct4-GFP (left panel) was labeled with PE-anti-TMEM-98, -GPR64 and -FAT4 and analyzed by flow cytometry based on the relative GFP levels (n = 5; G and H). Further assessment for TMEM-98, GPR64 and FAT4 within Oct4hi BCCs was based on size. Oct-4-GFP transfected MDA-MB-231 cells were gated based on GFP expression (I, top second panel) followed by gating of the 5% of cells with the highest GFP (I, R3, top third panel-Oct4hi). R3 was divided based on side (SSC) and forward scatter (FSC) (R4-R6) and then analyzed for TMEM98, GPR64 and FAT4 (I, lower panels). The results represent four independent experiments.
