Fig. 5: HMG20A is essential for cardiomyocyte and neural crest differentiation in mESCs.

A Validation of three Hmg20a DP clones by RT-qPCR. Shown is Hmg20a expression normalized to Hprt. Data is presented as mean ± SEM of three technical replicates. B Immunoblot analyses of endogenous HMG20A protein of extracts from WT and three Hmg20a DP mESC clones. H3 served as loading control (shown is one representative blot of two consistent replicates). C mESC neural crest cell (NCC) differentiation scheme. Created with BioRender.com. D RT-qPCR of neural crest and EMT marker genes in WT and three individual Hmg20a DP clones at Day9 of neural crest differentiation protocol normalized to Hprt, 18 S RNA and Gapdh expression. Data is presented as mean ± SEM of three technical replicates. E RT-qPCR of Hmg20a mRNA in WT and three individual Hmg20a DP clones at Day9 of neural crest differentiation protocol. Expression was normalized to Hprt, 18 S RNA and Gapdh expression. Data is presented as mean ± SEM of three technical replicates. F mESC cardiomyocyte (CM) differentiation scheme. Created with BioRender.com. G Depiction of percent beating (gray) or non-beating (black) WT or three individual Hmg20a DP EBs at Day7 (Top) or Day10 (bottom) of the cardiomyocyte differentiation procedure (see G). Ability to form contracting cardiomyocytes on Day7.5 is significantly reduced in all Hmg20a DP clones (Fischer’s exact test, two-sided, p = 3.1213974265e-087 (#06), p = 1.1876278705e-122 (#26), p = 2.0898552635e-091 (#48)). H RT-qPCR of Hmg20a mRNA in WT and three individual Hmg20a DP clones at Day7.5 of CM protocol. Expression was normalized to Hprt, 18 S RNA and Gapdh expression. Data is presented as mean ± SEM of three technical replicates. Source data for these figures are provided as a Source Data file.