Fig. 6

Evaluation of extracardiac Bmi1 ⁺ cells for de novo YFP⁺ CM generation. a Scheme for BM (RFP⁺) transplant into a lethally irradiated, TM-induced Bmi1-YFP mouse. b-e FACS analysis of the non-CM fraction of Bmi1-YFP mice after BM transplant (n = 3). b No YFP⁺ or RFP⁺ heart cells were detected in non-induced Bmi1-YFP mice. c FACS analysis of induced chimeric Bmi1-YFP hearts two months post-BM transplant confirmed the presence of YFP⁺ (3.53 ± 0.87 %) and RFP⁺ cells (45.5 ± 15 %). d Negative control for SCA-1 and CD45 staining of the non-CM fraction of Bmi1-YFP hearts (left) and percentage of SCA-1⁺CD45⁻ cells after the staining (20 ± 3.74 %; right). e Percentage of SCA-1⁺CD45⁻ in RFP⁺ cells in the non-CM fraction of chimeric Bmi1-YFP hearts two months post-transplant (0.26 ± 0.06 %). (f-h) FACS and ICC of the CM compartment of chimeric Bmi1-YFP mice two months post-transplant (n = 3). f No RFP or YFP expression was detected in Bmi1-YFP CM analyzed by FACS 5d-postTM (left). RFP⁺ CM from Act-RFP mice (right). g No RFP⁺ CM were detected in chimeric Bmi1-YFP mice two months post-transplant. h ICC of chimeric Bmi1-YFP CM two months post-transplant. Bars, 100 μm. Data shown as mean ± SD. YFP yellow fluorescent protein, CM cardiomyocytes, FACS fluorescence-activated cell sorting