Anti tom20 antibody sc 17764

For immunofluorescence assays, cells were fixed with 4% paraformaldehyde (PFA) and permeabilized with 0.2% Triton X-100. After blocking with 1% bovine serum albumin (BSA), cells were then incubated with the primary anti-calnexin antibody (AF18, 1:500, Thermo Fisher Scientific, United States), anti-TOM20 antibody (sc-17764, 1:500, Santa Cruz Biotechnology Inc., United States), anti-microtubule-associated protein 1A/1 B-light chain 3 (LC3B) (ab192890, 1:500, Abcam, United States) for 60 min at 37°C, respectively, followed by the corresponding secondary antibodies, Alexa 488 immunoglobulin G (IgG) (A32723, 1:1,000) or Alexa 568 IgG (A11011, 1:1,000) (Thermo Fisher, United States) in a dark chamber. Cells were examined using a laser scanning confocal microscope (LSM510, Carl Zeiss, Germany). Fluorescent images were acquired by a Plan-Neofluar 20×/0.40 LD or Plan-Apochromat 63×/1.40 Oil objective with either 488 nm laser excitation (525/50 nm band pass emission) or 543 nm laser excitation (570 nm long pass emission).

A Dual-Luciferase Reporter Assay kit (E1910 and E1960) was obtained from Promega Corporation. STZ (S01130) was purchased from Sigma-Aldrich. The anti-TOM20 antibody (sc-17764) and the anti-VDAC1 antibody (sc-390996) were purchased from Santa Cruz Biotechnology. Paricalcitol was provided by Professor Yanchun Li of the University of Chicago. The anti-collagen Ⅰ antibody (ab6308), anti-fibronectin antibody (ab2413), anti-PINK1 antibody (ab23707), anti-Parkin antibody (ab77924), anti-BNIP3 antibody (ab109362), and anti-VDR antibody (ab134826) were purchased from Abcam (Cambridge, UK). The anti-α-SMA antibody (19245) was purchased from Cell Signaling Technology.