Western blot (WB)

Total proteins were extracted from OPM-2, ANBL-6, KAS-6/1, and U266 cells using RIPA buffer (#9806, Cell Signaling Technology, Danvers, MA, USA). Equal amounts of protein were loaded and separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto a polyvinylidene fluoride (PVDF) membrane (#IPVH00010, Millipore, Billerica, MA, USA). Subsequently, the membranes were blocked with 5% non-fat milk for 1 h at room temperature (RT, 25℃) and then incubated with the primary antibody overnight at 4 °C. The following day, the membranes were washed with Tris-buffered saline containing 0.1% Tween 20 (TBST) and incubated with the corresponding secondary antibody at RT for 1 h. Finally, signals of the target proteins were detected using a Chemiluminescence Detection Kit (#P0018F, Beyotime Biotechnology). The primary antibodies used in this study included MUC20 antibody (1:300, #13380-1-AP, Proteintech, Rosemont, IL, USA), CDKN2A antibody (1:500, #ab270058, Abcam, Cambridge, UK), Met antibody (1:500, #ab216330, Abcam), Met (phospho Y1234 + Y1235) antibody (1:500, #ab278552, Abcam), CDKN2A (phospho Ser326) antibody (1:300, #GTX32378, GeneTex, Hsinchu City, Taiwan, China), ferredoxin 1 (FDX1) antibody (1:1000, #12592-1-AP, Proteintech), lipoyl synthase (LIAS) antibody (1:1000, #11577-1-AP, Proteintech), dihydrolipoamide S-acetyltransferase (DLAT) antibody (1:2000, #13426-1-AP, Proteintech), Metal response element binding transcription factor 1 (MTF1) antibody (1:500, #25383-1-AP, Proteintech), insulin-like growth factor receptor-1 (IGF-1R) antibody (1:1000, #ab182408, Abcam), His antibody (1:1000, #66005-1-Ig, Proteintech), kinesin family member 3 C (KIF3C) antibody (1:500, #ab236748, Abcam), and GAPDH (1:10,000, #KC-5G5, Aksomicks, Shanghai, China).