Fig. 1. Generation and identification of VDAC1^-/- H9c2 cells. (A) Rat VDAC1 genomic graph and the sites and sequences of two single guide (sg) RNAs in the rat genome. (B) Diagram of complex formation of sgRNA, T7 promoter, CRISPR guide RNA and Cas9 protein. (C) The gene mutation sequences in selected VDAC1^-/- H9c2 cell clones from sgRNA1 and 2. (D) Alignment of DNA sequencing traces of VDAC1^-/-C3 to WT H9c2 cells. Arrow in the box indicates a single nucleotide T deletion in the VDAC1^-/-C3 H9c2 cell clone. (E) Western blot analyses of selected WT and VDAC1^-/- H9c2 cell clones with VDAC1 specific and Tom-20 (loading control) antibodies. (F) Western blot analyses of selected WT and VDAC1^-/- H9c2 cell clones with VDAC2, VDAC3 and β-tubulin (loading control) antibodies.