Mouse anti kras

The cultured cells were washed with cold PBS and then treated with cell lysis buffer (2× loading buffer, 2 μg·mL⁻¹ Aprotinin, 1 mm PMSF, 2 mm β‐mercaptoethanol) at 100 °C for 10 min. Then the mixture was centrifuged under 4 °C at 14000g for 10 min. Approximately 10 μL of protein was loaded in each lane of 10% SDS/PAGE gel and separated, and the protein then transferred to a PVDF membrane. The membrane was blocked with 5% BSA for 1 h at room temperature and then incubated with primary antibodies at 4 °C overnight, followed by the secondary antibody. The antibodies used were rabbit anti‐CD133 (Cell Signaling Technology, Danvers, MA, USA; #64326), rabbit anti‐OCT4 (Cell Signaling Technology; #2750), rabbit anti‐SOX2 (Cell Signaling Technology; #3579), rabbit anti‐NANOG (Cell Signaling Technology; #4903), mouse anti‐KRAS (Santa Cruz Biotechnology, Dallas, TX, USA; sc30), mouse anti‐β‐Tubulin (Cell Signaling Technology; #6181), rabbit anti‐TET3 (Cell Signaling Technology; #85016), rabbit anti‐Lin28B (Signalway Antibody LLC, College Park, MD, USA；#21626), rabbit anti‐PKCβ (Cell Signaling Technology; #46809), rabbit anti‐PKCδ (Cell Signaling Technology; #2058), rabbit anti‐PKCζ (Cell Signaling Technology; #9372), rabbit anti‐PKCμ (Cell Signaling Technology; #2056) and mouse anti‐Flag (Sigma; CAT F1804).