Gapdh

Antibodies for western blot analysis were as follows: MACC1 (Abcam, Cambridge, MA, USA), Met (Cell Signaling Technology, Danvers, MA, USA), AKT (Cell Signaling), p‐AKT (Ser473) (Cell Signaling), E‐cadherin (Abcam), N‐cadherin (Abcam), vimentin (Abcam) and GAPDH (G8140, US Biological, Swampscott, MA, USA). The goat anti‐rabbit/mouse secondary antibodies were obtained from Cell Signaling. Two days post‐transfection, total proteins were extracted, followed by quantification with a Pierce BCA protein assay kit (Pierce, Bonn, Germany). Proteins were separated by 10% SDS‐PAGE gels and transferred to poly(vinylidene difluoride) membranes. After blocking, blots were probed with specific antibodies.

The whole proteins were lysed in RIPA buffer supplemented with protease and phosphatase inhibitors (Roche) and the concentrations were quantified with BCA Protein Assay Kit (Tiangen, Beijing, China), and an equal amount of 40 μg protein was separated by 10% SDS-PAGE gel and then transferred onto PVDF membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 5% nonfat milk in TBST for 2 h at room temperature and incubated overnight with specific primary antibodies at 4 °C. Then the membranes were washed three times by TBST and incubated with HRP-conjugated secondary antibody for 2 h at room temperature (ZSGB-BIO, China). Detection was performed by enhanced chemiluminescence kit (Amersham, Little Chalfont, UK). GAPDH (G8140; US Biological, Swampscott, MA, USA) was used as protein loading control. The SRPK1 primary antibody was obtained from Abcam (Cambridge, MA, USA). The antibodies against E-cadherin, N-cadherin, Vimentin, AKT, p-AKT, ZO-1, ZEB1, Slug, Snail, Twist, ERBB2, CCND1 and MCM2 were purchased from Cell Signaling Technology (Beverly, MA, USA).

The primary antibodies used in the immunoblotting assays were: SNAI1 (ab117866; Abcam) and GAPDH (G8140; United States Biological, Swampscott, MA, USA). Horseradish peroxidase-conjugated secondary antibody (Bio-Rad, Hercules, CA, USA) was used at a 1:1,000–1:5,000 dilution and detected using a Western Blotting Luminol Reagent (sc-2048; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), as described in a previous study (22 (link)).

Antibodies to STAT6 (D3H4, Cell Signaling technology), p-STAT6 (Tyr-641, Cell Signaling Technology), FLAG (M2, Sigma), p53 (Do-1, Santa cruz), PARP1 (F2, Santa cruz), Caspase-3 (H-277, Santa cruz), TRIML2 (ARP55636-P050, Aviva System Biology), EBV ZEBRA (sc-53904, Santa cruz), ICP0 (11060, Santa cruz), Ub-K48 (05–1307, EMD Millipore),Ub-K63 (05–1308, EMD Millipore), LC3B (GT3612,GeneTex) and GAPDH (G8140-01, US Biological) were used according to the manufacturers specifications. The monoclonal antibody anti-myc (9E10) and HA (12CA5) were prepared from hybridoma cultures. Mouse monoclonal antibodies against RTA were kindly provided by Ke Lan from Shanghai Pasteur Institute of CAS. Antibodies against IE1/2 were kindly provided by Zhikang Qian from Shanghai Pasteur Institute of CAS. Tetradecanoyl Phorbol Acetate (TPA) was purchased from Sigma and sodium butyrate from J&K Corporation. Proteasome inhibitor MG132 was purchased from Biomol International. 3-Methyladenine (3-MA), Choloquine (Chl), Cyclohexamide (CHX) and Hygromycin B were purchased from Sigma–Aldrich. Doxycycline (DOX) was purchased from Sangon Biotech (Shanghai). PMSF, Leupeptin, Aprotinin, Pepstatin A and Puromycin were purchased from Amresco, and G418 from Inalco S.p.A.