FIGURE 2:

The S2 site encoded by Opa1 exon 5b is necessary for OXPHOS-induced fusion in vivo. (A) Schematic of exon 5b, showing locations of S2 and the CRISPR-Cas9 gRNA target. Deletions present in clones 1 and 2 are indicated. (B) Schematic of OPA1 isoform composition after exon 5b–containing isoforms are eliminated; Red Xs indicate isoforms expected to be missing in mutant cells. (C) Western blot analysis of ∆exon 5b MEF clones. Clones 1 and 2 are positive clones that show the expected disappearance of bands a, c, and d. The last lane is a negative clone. Tubulin was used as loading control. (D) Quantification of mitochondrial morphology in WT and two ∆exon 5b clones in the indicated media. One hundred cells were counted for each experiment. Error bars indicate SD from three independent experiments.