Fig. 8
Generation of HSPB7 mutant hESCs. (A) Structure and partial sequence of the human HSPB7 gene, showing CRISPR targeting immediately downstream of the start codon and resultant HSPB7 homozygous and compound heterozygous mutant clones that were recovered. (B) qPCR quantification shows expression levels of pluripotency markers are normal in HSPB7 mutant hESC clones. HSPB7 mutant lines are competent to generate cardiomyocytes, and do not express HSPB7 protein as shown by flow cytometry (C,D) and immunofluorescence (E), both performed on day 10 of differentiation. (F) qPCR quantification of cardiac HSPB family members in wildtype and HSPB7 mutant hESC-derived cardiomyocytes. Changes in gene expression are significant according to an unpaired t-test, corrected for multiple comparisons with the Holm-Sidak method.
Reprinted from Developmental Biology, 435(1), Mercer, E.J., Lin, Y.F., Cohen-Gould, L., Evans, T., Hspb7 is a Cardioprotective Chaperone Facilitating Sarcomeric Proteostasis, 41-55, Copyright (2018) with permission from Elsevier. Full text @ Dev. Biol.