Fig. 7

Fig. 7

Essentiality of UGP2 and other disease candidate genes with a similar mutation mechanism. a qRT-PCR analysis of the hematopoietic stem cell markers GATA2, LMO2 and RUNX1, after 12 days of differentiation of wild type, UGP2 KO and UGP2 KO rescue ESCs. Shown is the mean fold change for the indicated genes compared to wild type, normalized for the housekeeping gene TBP. Results of two biological and two technical replicates are plotted. Error bars represent SEM; *p < 0.05; **p < 0.01, ***p < 0.001, unpaired t test, two tailed. b As a, but now for cardiomyocyte differentiation at day 15, assessing expression of the cardiomyocyte markers TNNT2, MYL2 and MYL7. c Bright-field image of cardiomyocyte cultures of wild type, UGP2 KO and rescue cells. Note the elongated organized monolayer structure cardiomyocytes capable of beating in wild type and rescue cells that are absent in KO cultures. Scale bar is 400 µm. d Scheme showing the homology search to identify genes with a similar structure as UGP2, where ATG-altering mutations could affect a tissue-specific isoform causing genetic disease. e Heat map showing the ratio of short isoform expression over total isoform expression from published RNA-seq data amongst 20 tissues for 83 out 247 essential genes that are not yet implicated in disease and in which the short and longer protein isoforms differ by less than 50 amino acids at the N-terminal