Ab206423

The cell lines HEK293T were got from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cell lines KYSE30 and OE19 were obtained from fenghbio (Hunan, China). Antibodies for ATF5 (ab184923), VEGFA (ab52917), TIMP1 (ab61224), TWIST1 (ab175430), EGF (ab206423), CD31 (ab134168) and POL II (ab264350) were bought from Abcam (Cambridge, MA, USA). Antibodies for PDK1 (13037), PGK1 (68540), CA9 (5649), CBP (7389), P300 (54062), HIF1α (14179), HIF1β (5537) and flag-tag (2368) were procured from Cell Signaling Technology. The human VEGFA ELISA Kit (EK0539) was purchased from BOSTER Biological Technology (Wuhan, China). Identified primers were synthesized by TSINGKE Biological Technology (Beijing, China). For hypoxic conditions, cells were cultured in a sealed hypoxia chamber with a mixture of 1% O2, 5% CO2, and 94% N2 for 2 h.

The following antibodies were used for western blot: MYL1 (Abcam, ab151749, 1:1000), EGF (Abcam, ab206423, 1:1000), EGFR (Abcam, EP38Y, 1:1000), β-actin (Abcam, ab8226, 1:1000), Goat anti-mouse second antibody (Cell Signaling Technology, 5470 S, 1:15000), Goat anti-rabbit second antibody (Cell Signaling Technology, 5151 S, 1:30000). PENTER control plasmid (Shandong Vigene Biosciences, Jinan, China) and pENTER-MYL1 plasmid (Shandong Vigene Biosciences, Jinan, China) were used for transfection.