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Fig. 4 | Cell & Bioscience

Fig. 4

From: BVES is a novel interactor of ANO5 and regulates myoblast differentiation

Fig. 4

Generation of Ano5-KO and BVES-KO C2C12 cells by gene editing. A Design of two TALEN pairs (E5A and E5B) targeting the exon 5 of mouse Ano5. The left and the right target sites were colored in red, with the repeat-variable di-residues (RVD) of each TAL shown below. B T7E1 assay of the gene editing activity conferred by E5A and E5B TALEN pairs in C2C12 myoblasts. The red arrows indicate the T7E1 enzyme cleavage bands shown in TALEN-transfected cells but not in control (Ctrl) samples. C Screening of individual C2C12 clones transfected with E5A TALEN pair by T7E1 assay. The blue arrows indicate the predicted T7E1 cleavage products. The red arrow indicates a smaller product appeared in Clone 8. D RT-PCR analysis of Ano5 expression in control and Clone E5A8-12. E Generation of Bves-KO C2C12 cells by CBE base editing. The gRNA targeting the exon 4 of mouse Bves was designed to install a premature TAG stop codon at position 159 (highlighted in red). Sanger sequencing confirmed the genotype of BVES-KO C2C12 cells. F Western blot analysis of BVES expression in the total membrane fractions of WT, BVES-KO and Ano5-KO C2C12 myoblasts

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