CsGRN (5 μg) and CsESPs (30 μg) were subjected to 12% SDS-PAGE and then were electro-transferred onto polyvinylidene difluoride (PVDF) membranes (Whatman, Maidstone, UK) at 100 V for 1 h in a Trans-Blot transfer (Bio-Rad). The membranes were blocked with 5% (w/v) skimmed milk in PBS for 2 h at room temperature and then were probed with mouse anti-His tag monoclonal antibody (1:2000, Novagen), rat anti-CsGRN serum (1:200), rat anti-CsESPs serum (1:200) and rat naïve serum (1:200) at 4 °C overnight, respectively. After washing with PBS, the membranes were successively incubated with HRP-conjugated rabbit anti-rat or rabbit anti-mouse IgG at 1:2000 dilution (Proteintech Group) at room temperature for 1 h. The blots were visualised by enhanced chemiluminescence (ECL, Millipore, Billerica, USA). We also used Western blotting to detect the mechanism of cancer progression induced by CsGRN. In detail, the total proteins of transfection cells were extracted using radioimmunoprecipitation assay lysis buffer (RIPA, Beyotime, Shanghai, China) and then were electrotransferred onto PVDF membranes. The membranes were incubated at 4 °C overnight with E-cadherin (1:2,000), vimentin (1:2,000), N-cadherin (1:2,000), ZO-1 (1:2,000), β-catenin (1:2,000), or GAPDH (1:2,000) diluted in blocking solution. In addition, the membranes with total proteins from Balb/c mice liver tissues were incubated at 4 °C overnight with E-cadherin (1:2,000), vimentin (1:2,000), N-cadherin (1:2,000), ZO-1 (1:2,000), β-catenin (1:2,000), p-ERK (1:2,000), ERK (1:2,000), p-AKT (1:2,000) or GAPDH (1:2,000) diluted in blocking solution, all antibody were products from cell signaling technology (CST, Boston, USA).