Anti c jun

Western blotting was performed as previously described,^{35 (link)} using anti-CDK5, anti-FAK, anti-p-FAK (Ser732), anti-PAK1, anti-p-PAK1 (Thr212), anti-ERK5, anti-p-ERK5 (Thr218/Tyr220) (Abcam), anti-p-ERK5 (Thr732), anti-p35 (Cell Signaling Technology, Danvers, MA, USA; anti-p-ERK5 Thr732 was custom-made from CST), anti-c-fos, anti-c-jun (Bioworld Technology, Louis Park, MN, USA), anti-c-myc, anti-VEGFA and anti-MMP1 antibodies (Proteintech, Chicago, IL, USA). Loading control was used with a mouse anti-β-Actin monoclonal antibody (Proteintech).

Cells were lysed and homogenized in RIPA buffer supplemented with complete protease inhibitors. Equal amount of proteins (200 µg) was resolved in 12% SDS-PAGE. The proteins were then transferred onto PVDF membranes following electrophoresis. After blocked in 5% (w/v) non-fat milk for 1 h at room temperature, the membranes were then incubated at their respective appropriate dilutions of specific primary antibodies overnight at 4°C. The sources of primary antibodies were: anti-Oct4, anti-Sox2, anti-Vimentin and anti-phospho-STAT3 (Signalway Antibody, USA), anti-GAPDH (Kangcheng, Shanghai, China), anti-E-cadherin, anti-ERK1/2, anti- phospho-ERK1/2 and anti-N-cadherin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-PCNA, anti-STAT3, anti-NF-κB, anti-phospho-NF-κB, anti-CyclinD1, anti-VEGF and anti-C-Jun (Bioworld Technology, Louis Park, MN, USA), anti-C-myc (ProteinTech Group, Chicago, IL, USA).