Rabbit anti cyclin d2

Total protein from cells was extracted using RIPA lysis buffer (CW2333, CWBIO, Beijing, China) supplemented with 1 mmol/L phenylmethylsulfonylfluoride (A32961, Thermo Fisher, MA, USA), followed by separation with 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transfer onto PVDF membranes. The membranes were then blocked in TBST containing 5% milk for 1 h at room temperature and incubated at 4 °C overnight with rabbit anti-TGFBI (2719S, Cell Signaling Technology, MA, USA) and rabbit anti-cyclin D2 (3741, Cell Signaling Technology, MA, USA). Proteins were visualized using HRP-conjugated secondary antibodies. The bands were exposed by an ECL Kit as previously described. (17001102, Millipore, MA, USA).

Whole lysates of cells were prepared as previously described. Western blotting was performed with rabbit-anti-cyclin D2 (Cell Signaling Technology), mouse-anti-cyclin E (Cell Signaling Technology), rabbit-cyclin A2 (NOVUS Biologicals), mouse-cyclin D3 (Cell Signaling Technology), mouse-anti-β actin (Santa Cruz Biotechnology), rabbit-anti-ErbB-1 (Santa Cruz Biotechnology), rabbit-anti-ErbB-2 (Santa Cruz Biotechnology), rabbit-anti-ErbB-3 (Santa Cruz Biotechnology), mouse-anti-ErbB-4 (Santa Cruz Biotechnology), mouse-anti-phospho-Tyr (Millipore), rabbit-anti-phospho-Erk1/2 (Cell Signaling Technology), rabbit-anti-Erk1/2 (Cell Signaling Technology), rabbit-anti-phospho-Akt (Ser473) (Cell Signaling Technology), rabbit-anti-Akt (Cell Signaling Technology), rabbit-anti-phospho-mTOR (Cell Signaling Technology), rabbit-anti-mTOR (Cell Signaling Technology), or mouse-anti-GAPDH (Santa Cruz Biotechnology).