Clone 18

Cells were harvested, and total cell protein lysates were prepared as previously described (Xu et al., 2011 (link)). In brief, cells were collected, washed with ice-cold PBS, resuspended in SDS PAGE sample buffer, and incubated at 100°C for 10 min. Protein concentrations were measured directly in the samples using NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific). Proteins were separated on 4–20% gradient Tris-Glycine polyacrylamide gels (Invitrogen), transferred onto PVDF membranes (Immubilon-P; Millipore), and incubated with primary antibodies, followed by HRP-conjugated anti–mouse (Roche) or anti–rabbit (Roche) secondary antibodies. Blots were detected with ECL PLUS reagent (GE Healthcare) using ImageQuant LAS4000 chemiluminescence imager (GE Healthcare). Antibodies used for Western blot were: anti-cortactin mAb (clone 4F11; Millipore), anti-cofilin polyclonal antibody (clone FL-166; Santa Cruz Biotechnology, Inc.), anti-Arp2 polyclonal antibody (clone H-84; Santa Cruz Biotechnology, Inc.), anti-ARC/p34 polyclonal antibody (Millipore), anti-Tom20 polyclonal antibody (clone FL-145; Santa Cruz Biotechnology, Inc.), anti-Opa1 mAb (clone 18; BD), and anti-Mfn2 mAb (Abcam).