Ab37155

The protein extracted from HCC Cells or tissues were lysed in ice-cold radioimmunoprecipitation assay buffer and then incubated with primary antibodies overnight at 4°C. The primary antibodies are as follows: Tollip (1:1,000, ab37155, Abcam), Total-AKT (1:1,000, 4691, Cell Signaling Technology), p-AKT (1:1,000, 4046, Cell Signaling Technology), Total-mTOR (1:1,000, BS1555, Bioworld Technology), p-mTOR (1:1,000, BS470, Bioworld Technology), E-Cadherin (1:1,000, ab231303, Abcam), N-Cadherin (1:1,000, ab76011, Abcam), Snail (1:1,000, ab180714, Abcam), Vimentin (1:1,000, ab92547, Abcam), and GAPDH (1:1,000, SC-25778, Santa Cruz). The relative expression was obtained using a FluorChem E Imager (ProteinSimple, FluorChem E), and membranes were treated with enhanced chemiluminescence reagents (170-5061; Bio-Rad). The results were shown after normalizing against GAPDH gene expression.

The HCC tissues and cell sections were prepared according to the protocols. Following incubation with primary antibodies overnight at 4°C, the slides were washed with phosphate-buffered solution and incubated with the relevant secondary antibodies. The primary antibodies are as follows: Tollip (ab37155, Abcam) and HNF4 (ab41898, Abcam). The images were analyzed using Image-Pro Plus 6.0.