Fig. 7

Expression analysis of potassium pathway-related genes and proteins in iPSC-CMs. A Venn diagram showing differentially expressed genes between the two groups at the same stages. B PCA scores from proteome-seq data for late iPSC-CMs (day 30) in two groups. Blue, CRISPR group; orange, mutation group. Heatmap analysis for the transcriptome (C) and proteome (D) (Mutation n = 3, CRISPR n = 3). E Comparison for the overall regulatory status of potassium channel-related pathways (including the participation of KCNJ2) at the gene expression level (upper row) and protein expression level (lower row) in the two groups (Mutation n = 3, CRISPR n = 3). F The overall regulatory status of four potassium ion-related pathways in CMs of the mutant group were significantly downregulated after comparison (Mutation n = 3, CRISPR n = 3). G A total of 3 target genes and their protein (CTTN, KCNJ2, ATP1B1) expression were consistently decreased in the mutant group by pathway expression analysis in a Venn diagram. H qRT‒PCR was performed to assess the expression of the target genes KCNJ2, ATP1B1, CTTN, and ZNF528, with β-actin used as a housekeeping gene (Mutation n = 6, CRISPR n = 6). I, J Detection of target proteins in iPSC-CMs (day 30) of the three groups (mutation, CRISPR, healthy control) by Western blotting (Mutation n = 3, CRISPR n = 3, Control n = 3). β-Actin was used as a loading control. Bands were quantified with Image J software. One asterisk indicates p < 0.05, and two asterisks indicates p < 0.001 compared with the other groups. Data are shown as mean ± SD.*p < 0.05; data were compared by two-sample t-test