β actin antibody ap0060

The total protein of cells was prepared as previously described [32 (link)]. Sample proteins (30 μg) were separated via SDS-PAGE, transferred to nitrocellulose membranes, and blocked with 5% non-fat milk. After incubation with the indicated antibodies, the membranes were washed and incubated with the secondary antibodies. Immunoreactivity was detected using ECL reagents. The following primary antibodies were used in this study: β-actin antibody (AP0060, Bioworld, Nanjing, China), TLR4 antibody (sc-293072, Santa Cruz Biotechnology, Paso Robles, CA, USA), Cav-1 antibody (ab192869, ABCAM, Cambridge, UK), Ras homolog family member A (RhoA) antibody (2117S, Cell Signaling Technology, Danvers, MA, USA), p65 antibody (10745-1-AP, Cell Signaling Technology), p-p65 antibody (3033S, Cell Signaling Technology), IκBα antibody (4814S, Cell Signaling Technology), and p-IκBα antibody (2859S, Cell Signaling Technology).

GANT61 purchased from Sigma-Aldrich (St. Louis, MO, USA) was dissolved in DMSO and stored at −20 °C. Cisplatin was obtained from Sigma-Aldrich and was dissolved at a stock concentration of 2 mmol/L. Rhodamine 123 was purchased from Sigma-Aldrich. Foetal bovine serum (FBS) and 0.25 % trypsin/EDTA were bought from Gibco Life Technologies (Carlsbad, CA, USA). A FITC-annexin V kit was purchased from Abcam (Cambridge, MA, USA). The CCK-8 cell count kit for cell proliferation analysis was purchased from Tongren Chemical Research Institute (Kyushu, Japan). The reverse transcription kit was purchased from Takara (Shiga, Japan). SYBR Green I was purchased from Noble Ryder (Beijing, China). Antibodies for GLI1 (ab49314), GLI2 (ab26056), Bax (ab10813), BCL-2 (ab59348), caspase-3 (ab32351) and caspase-9 (ab25758) were purchased from Abcam (Cambridge, MA, USA). The β-actin antibody (AP0060) was purchased from Bioworld (Louis Park, MN, USA).