Goat anti tfam

Purified lung epithelial cells were lysed in 2x Western blotting buffer (120 mM Tris-Cl pH 6.8, 4% SDS, 20% Glycerol, 200 mM DTT, 0.02% Bromophenol Blue). The lysates were centrifuged at 16,100 x g for 15 min at 4 °C before loading onto SDS-PAGE. The primary antibodies used were rabbit anti-Raptor (1:2000, EMD Millipore Corporation, Cat# 09-217; RRID:AB_1659713), mouse anti-S6 ribosomal protein (1:2000, Cell Signaling Technology, Cat# 2317; RRID:AB_2238583), rabbit anti-phospho-S6 ribosomal protein (Ser235/236) (1:2000, Cell Signaling Technology, Cat# 4856), rabbit anti-MLC2 (1:500; Cell Signaling Technology, Cat#3672; RRID:AB_10692513), goat anti-TFAM (1: 250, Santa Cruz Biotechnology, Cat# sc-23588; RRID:AB_2303230), rabbit anti-COX10 (1:500, Proteintech, Cat# 10611-2-AP; RRID:AB_2084833), mouse anti-MTCO1 (1:500, Abcam, Cat# ab14705; RRID:AB_2084810), and mouse anti-alpha-tubulin (1:3000, Developmental Studies Hybridoma Bank, Cat# 12G10; RRID:AB_1157911). Western blotting images were captured on a LI-COR Odyssey Fc device. The uncropped images are available in the Source Data file.

The following primary antibodies were used in immunoblotting: rabbit anti‐NEK5 (Atlas Antibodies®, HPA, HPA035565, 1 : 1000); rabbit anti‐LonP1 (Sigma‐Aldrich, Atlas Antibodies®, HPA, HPA002034); mouse anti‐Nek5 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA, G‐12, sc‐515457); mouse anti‐GAPDH (Santa Cruz Biotechnology, FL‐335, sc‐25778, 1 : 1000); goat anti‐TFAM (Santa Cruz Biotechnology, E‐16, sc‐30963‐1 : 500); mouse anti‐tubulin (Santa Cruz Biotechnology, Tu‐02, sc‐8035, 1 : 1000); mouse anti‐Tom20 (BD Biosciences, Franklin Lakes, NJ, USA, 612278, 1 : 1000); mouse anti‐FLAG M2 (Merck, 1 : 5000); and Myc‐Tag (9B11) mouse mAb (Cell Signalling Technology, Danvers, MA, USA #2276 1 : 5000).
For immunoprecipitation (IP), anti‐FLAG® M2 Affinity Gel (Millipore, Billerica, MA, USA A2220) was used.
For immunofluorescence, rabbit anti‐LonP1 (Atlas Antibodies®, HPA, HPA002034, 1 : 100); mouse anti‐Nek5 antibody (Santa Cruz Biotechnology, G‐12, sc‐515457, 1 : 100); and Alexa Fluor‐conjugated secondary antibodies were used at 1 : 500 dilution: anti‐goat Alexa Fluor 647; anti‐mouse Alexa Fluor 488; and anti‐goat Alexa Fluor 488. MitoTracker™ Deep Red FM (Invitrogen, M22426) was used for mitochondrial visualization.