Fig. 6

HAND1/2 modulated cardiomyocyte differentiation through TBX5. A Venn diagram of HAND1 and HAND2 ChIP-seq and downregulated DEGs in H1/H2-dKO cells compared to WT cells in RNA-seq. B Expression of some TFs in WT and H1/H2-dKO sorted NKX2.5-eGFP⁺ cells at differentiation day 7. Relative to GAPDH expression (n = 3). Unpaired t test. C The ratio of Luciferase and Renilla activity in HAND1 and HAND2 overexpression 293T cells (n = 6). CTRL represented 293T transfected with pGL3-TBX5-Luciferase and Renilla only. One-way ANOVA. D TBX5 overexpression after DOX induction verified by western blot at differentiation days 5 and 10. GAPDH served as loading control. Corresponding uncropped full-length gels and blots are presented in Additional file 8: Fig. S11. E, F The percentage of NKX2.5-eGFP⁺ cells in WT and TBX5-OE cells (−DOX and + DOX) at differentiation day 7 (n ≥ 3). One-way ANOVA. G Expression of NKX2.5, MYH6 and TNNT2 in WT and TBX5-OE cells (−DOX and + DOX) at differentiation day 7. Relative to GAPDH expression (n = 3). One-way ANOVA. H Expression of SHF markers in WT and TBX5-OE cells (−DOX and + DOX) at differentiation days 5 and 7. Relative to GAPDH expression (n = 3). One-way ANOVA. I Expression of atrial, OFT and ventricular cardiomyocyte markers in WT and TBX5-OE cells (−DOX and + DOX)-derived differentiation day 30 cardiomyocytes. Relative to GAPDH expression (n = 3). One-way ANOVA. J Comparison of corrected APD90, APD50 and APD90/APD50 ratio recorded by MEA in WT and TBX5-OE cells (−DOX and + DOX)-derived differentiation day 30 cardiomyocytes (n ≥ 8). One-way ANOVA. K Comparison of APD90 and APD50 recorded by whole-cell patch clamp in WT and TBX5-OE cells (−DOX and + DOX)-derived day 30 cardiomyocytes (n ≥ 9). One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. n.s: non-significant