Fig. 1

Characterization and evolution of B-CPC throughout Bmi1-YFP mouse lifespan. a Generation of Bmi1 ^CreER/+;Rosa26 ^YFP/+ (Bmi1-YFP) mice. b Detection of the YFP⁺ fraction (2.7 ± 0.2%, n = 21) of freshly isolated mononuclear non-cardiomyocyte heart cells from Bmi1-YFP mice, analyzed five days post TM induction (5d-postTM); inset, Bmi1-YFP^NI (non-induced) and TM-induced Rosa26 ^YFP/+ negative controls. Data shown as mean ± SEM. PE, phycoerythrin. c Immunofluorescence analysis of BMI1 and SCA-1 in freshly isolated YFP⁺ cells. Bars, 50 μm. d The B-CPC population is a subset of the SCA-1⁺ population (5.4 ± 0.4 %, n = 18). The plots show from (left to right and top up to bottom) the YFP⁺ fraction of Bmi1-YFP hearts 5d-postTM, the negative control from SCA-1 staining in the non-CM fraction, staining for SCA-1, and the fraction of the YFP⁺ SCA-1⁺ population. Data represented as mean ± SEM. PE, phycoerythrin, SSC, side scatter. e RT-qPCR of freshly sorted Bmi1 ⁺ (YFP⁺) and SCA-1⁺ YFP⁻ cells (n = three replicates; two to three mice per replicate). Data shown as mean ± SD. P values were calculated by paired Student’s t-test. * P < 0.05, ** P < 0.01. f Analysis of the YFP⁺ compartment in Bmi1-YFP mice at one year post-TM induction (1y-postTM) (left) (5.8 ± 0.74 %) and analysis of B-CPC (YFP⁺) in one-year-old mice 5d-postTM (right) (2.2 ± 0.22 %). PE, phycoerythrin, SSC, side scatter. g YFP⁺ cell number at 5d-postTM in young mice (two-month-old; n = 15), in mice 1y-postTM induction at six to eight weeks of age (n = 4), and in one-year-old mice at 5d-postTM (n = 7). Data shown as mean ± SEM. P values were calculated by unpaired Student’s t-test with Welch’s correction, compared to 5d-postTM young mice. * P < 0.05, *** P < 0.0001