Immunofluorescence analysis of DNA synthesis (BrdU assay) and cardiac Troponin I (cTnI) on neonatal (postnatal day 1, P1) cardiomyocytes following SMAD1, SMAD5, and SMAD9 knockdown, alone or in combination, upon BMP7 stimulation at 10 ng/mL for about 48 h (n= 19,167 cardiomyocytes pooled from the analysis of 54 samples); representative pictures are provided; scale bars, 20 m, arrows point at proliferating cardiomyocytes. The values in (A)–(E) are presented as mean (error bars show SEM); statistical significance was determined using one-way ANOVA followed by Sidak’s test (comparison between pairs of treatments) in (A), (B), (D), and (E); and using two-sided Student’s t test in (C).
