Gapdh antibody

GAPDH antibody was obtained from Sangon Biotech (Shanghai, China). E-cadherin and β-catenin antibodies were purchased from abcam (Shanghai, China). HIF-1a antibody was obtained from ABclonal (Wuhan, China). Alexa Fluor 488-conjugated secondary antibody was purchased from Thermo Fisher Scientific (Shanghai, China). House dust mite (HDM) was purchased from Greer (Beijing, China). Ammonium acetate (NH4AC) was purchased from Sigma Aldrich (St. Louis, MO, USA). Acetonitrile was purchased from Merck (Shanghai, China), and ammonium hydroxide (NH4OH) and methanol were purchased from Fisher.

After transfection of miRNA-122 mimic and miRNA-122 inhibitor groups, the total protein of HUVECs cells was extracted by adding RIPA buffer (Vazyme Biotech, China). Protein samples were separated by SDS-PAGE and transferred to polyvinylidene fluoride membrane by polyacrylamide sulfate gel electrophoresis. After the transfer, the membrane was sealed with 5% Bowene Serum Albumin at room temperature for 1h and subsequently incubated overnight with the required primary antibody at 4°C. The next day, the membrane was incubated with Sangon Biotech (China, 1:500 dilution) for 1h, and the protein was observed by enhanced chemiluminescence (Vazyme Biotech, China). G:BOX Chemi XX9 imaging system (Syngene, UK) for protein visualization was used. Individual western blotting was performed with primary antibody, Bax, Bcl-2, Caspase-3 antibody (Abcam, USA, 1:1000 diluted), Hes1, Notch1, Vascular Endothelial Growth Factors [VEGF], CCNG1 and GAPDH antibody (Sangon Biotech, China, 1:500 diluted).