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. Author manuscript; available in PMC: 2021 Apr 14.
Published in final edited form as: Circulation. 2020 Feb 11;141(15):1249–1265. doi: 10.1161/CIRCULATIONAHA.119.043067

Figure 6. Loss of function study of Pkm2 expression in CMs during heart development reduces heart size, increases CM size, lowers total CM number and limits CM cell division capacity.

Figure 6.

A. Experimental plan for generating inducible CMSWT (Pkm2+/+, (B6129SF2/J) littermates expressing Cre exclusively in CMs) or CMSKO-Pkm2 mice. B. Experimental timeline for evaluating the effect of CMSKO-Pkm2 on CM cell division and heart development. C. Representative images of E18 CMSWT or CMSKO-Pkm2 mice and evaluation of body weight (D. n=7). E. Representative images of Pkm2 and α-Actinin immunostaining as well as whole heart H&E staining F. Evaluation of heart size (G. n=4) and heart weight to body weight ratio (H. n=7) of E18 CMSWT or CMSKO-Pkm2. Representative images of WGA (I.) and quantification thereof (J. n=7). Quantification of CM numbers isolated from heart (K. n=4) or counted per section (L. n=7) in E18 CMSWT or CMSKO-Pkm2 mice. M. Representative images of pH3+ or Ki67+ CMs in E18 CMSWT or CMSKO-Pkm2 hearts. Quantification of pH3+ (N. n=7) or ki67+ (O. n=7) CMs in E18 CMSWT or CMSKO-Pkm2 hearts. P. Representative images of pH3- or Ki67-positive CMs in isolated CMs from E18 CMSWT or CMSKO-Pkm2 hearts. Quantification of pH3+ (Q. n=5) or ki67+ (R. n=5) CMs isolated from E18 CMSWT or CMSKO-Pkm2 hearts. S. Relative gene expression measured by qRT-PCR comparing E18 CMSWT or CMSKO-Pkm2 hearts (n=3). T. Representative images of Pkm1 expression in E18 CMSWT or CMSKO-Pkm2 hearts. White arrowheads point to CMs. Yellow arrowheads point to non-CMs. Unpaired two-tailed t-test for (D, G, H, J-L, N, O, Q, R and S). ****, P<0.0001, ***, P<0.001, **, P<0.01, *, P<0.05, N.S, Not Significant. Scale bar = 25μm (E and T), 5μm (I), 50μm (M&P).