Ab217673

Primary antibodies used for Western blot, immunohistochemistry and immunocytochemistry are listed as followed: Hif-1α (79233, Cell signaling, USA), α-SMA (ab32575, Abcam, USA), Bnip3 (ab10433, Abcam, USA; ab38621, Abcam, USA), LC3B (83506, cell signaling, USA; 12741, cell signaling, USA), vimentin (ServiceBio, GB12192, China), phosphor-vimentin Ser56 (ab217673, Abcam, USA), phosphor-vimentin Ser38 (ab52942, Abcam, USA), phosphor-vimentin Ser72 (ab52944, Abcam, USA), phosphor-vimentin Ser82 (ab52943, Abcam, USA), caspase 3 (ab197202, Abcam, USA), cytochrome C (sc-13156, Santa Cruz, USA), GAPDH (GB12002, Servicebio, China).

Cells were lysed for 10 min at 4°C with adding RIPA lysis buffer (Thermo Fisher Scientific). The solution was then put into a 4°C centrifuge at 12000 r/min for 15 minutes. The amount of protein loaded in each line was about 40 μg, and separated in the presence of 10% SDS-PAGE. Then, it was transferred to PVDF membrane. After that, the PVDF membrane was kept in 5% skimmed milk powder and held at room temperature for 1 hour. Then, the PVDF membrane was incubated overnight in the refrigerator at 4°C with primary antibody and incubated for 2 hours with secondary antibody. The antibodies were as follows: rabbit anti-P4HA3 (Biorbyt, orb77765), mouse anti-GAPDH (Abcam, ab9484), rabbit anti-E-cadherin (Abcam, ab18203), rabbit antivimentin (Abcam, ab217673), rabbit anti-N-cadherin (Abcam, ab76011), and rabbit antisnail (Abcam, ab216347).