Ab251807

Paraffin sections of xenograft tumor tissues were cut with a thickness of 4 μm. Sections were incubated with primary antibodies: anti-RTKN2 (Abcam, #ab251807, 1:1000), anti-β-catenin (Abcam, #ab16051, 1:1000), anti-Ki67 (Abcam, #ab245113, 1:1000), anti-E-cadherin (Abcam, #ab1416, 1:500), and anti-N-cadherin (Abcam, #ab76057, 1:1000). The temperature was maintained at 4 °C overnight. Sections were co-incubated with HRP‐polymer‐conjugated secondary antibodies after washing with phosphate‐buffered saline, then, they were immunostained using DAB plus kit.

Different groups of HCC cells were lysed on ice with RIPA buffer. Then, protein lysates were electrophoresed on 10% SDS polyacrylamide gels, transferred onto PVDF membranes (Millipore, USA), and blocked with 5% skim milk. Afterwards, primary antibodies: anti-RTKN2 (Abcam, #ab251807, 1:1000), anti-β-catenin (Abcam, #ab16051, 1:1000), anti-PCNA (Abcam, #ab29, 1:1000), anti-CyclinD1 (Abcam, #ab134175, 1:1000), anti-c-Myc (Abcam, # ab32072, 1:1000), anti-Bcl-2 (Abcam, # ab185002, 1:1000), anti-Bax (Abcam, # ab32503, 1:1000), anti-MMP-7 (Abcam, # ab207299, 1:1000), anti-E-cadherin (Abcam, #ab1416, 1:500), anti-N-cadherin (Abcam, #ab76057, 1:1000) were added at 4 °C overnight. Subsequently, secondary antibodies (1:1,000; Millipore, USA) was incubated for 1 h. Data was obtained and calculated using ImageJ software.