Anti vegfa ab1316

The uterine specimens collected at day 5 post-transplantation and hAECs exposed to H₂O₂ were lysed in cold RIPA buffer (Yeason, China) supplemented with protease inhibitor cocktail (Yeason, China) and phosphatase inhibitor cocktail (Yeason, China). The protein concentrations were then quantified by a Pierce BCA Protein Assay Kit (23225, Thermo Fisher Scientific, USA). Proteins were separated in polyacrylamide gel and transferred to the PVDF membranes. Membranes were blocked with 5% non-fat milk in Tris-HCI buffer solution containing 0.1% Tween-20 (TBST) and separately incubated with anti-MMP-8 (17874-1-AP, Proteintech, Chicago, USA), anti-VEGFA (ab1316, Abcam, USA), and anti-β-tubulin (30303ES10, Yeason, China) diluted with 5% BSA in TBST at 4 °C overnight. After washing with TBST, membranes were incubated with HRP-conjugated anti-rabbit IgG (Yeason, China). The blots were visualized with an Enhanced Chemiluminescence Kit (Millipore, USA). The level of β-tubulin was used as the internal standard. The immunoreactive band intensities were quantified by ImageJ software (NIH, MD, USA).