Western Blot Analysis of Cellular Proteins

At 48 h after transfection, protein samples were extracted from the cells and western blot analysis was performed following the protocols described previously (25 (link),26 (link)). The materials used for the analysis were as follows: RIPA buffer (Thermo Fisher Scientific, Inc.), Protease Inhibitor Cocktail (Sigma-Aldrich), DC Protein assay kit (Bio-Rad), polyacrylamide gels (Wako Pure Chemical), PVDF membrane (Bio-Rad), and PVDF Blocking Reagent for Can Get Signal^® (TOYOBO). The primary antibodies used were anti-GA (Abcam; EP7212), anti-LDHA (Abcam; EP1566Y), and anti-β-actin (Sigma-Aldrich; A2228). The secondary antibodies used were HRP-conjugated goat anti-rabbit and horse anti-mouse IgG (Cell Signaling Technology). The immunoblots were detected and visualized by Fusion-FX7 (Vilber Lourmat) with LuminataTM Forte Western HRP Substrate (Millipore). β-actin was used as an internal control.