Bs5085r

Tissues were fixed in 10% buffered formalin and embedded in paraffin. Paraffin sections of tumor tissues were prepared, dewaxed by dimethyl-benzene, and hydrated with different concentrations of ethanol (100%, 95%, 85%, 70%, and 50%). The following steps were performed: blocking of endogenous peroxidase activity in 3% H₂O₂ solution, unmasking of the antigenic epitope with citrate buffer, incubation with blocking buffer for blocking, and incubation with primary antibody (VEGF (Bioss, BS1665R), CD34 (Bioss, BS5085R), TGF-β1(Bioss, BS0086R), IL-10 (Bioss, BS6761R), and EGFR (Bioss, BS1007R)) and secondary antibody (horseradish peroxidase-labeled goat rabbit IgG (H + L) (Beyotime, A0208)). Lastly, the DAB (Beyotime) substrate solution was applied to reveal the color of antibody staining. Images were acquired by an Motic BA400 microscope (Motic, China). The expression of cytokines was analyzed by Image-Pro Plus 6.0 (Media Cybernetics, USA).

Tissues were fixed in 10% buffered formalin and embedded in paraffin. Paraffin sections of tumor tissues were prepared, dewaxed by dimethyl-benzene, and hydrated with different concentrations of ethanol (100%, 95%, 85%, 70%, and 50%). Subsequently, the following steps were performed: blocking of endogenous peroxidase activity in 3% H₂O₂ solution, unmasking of the antigenic epitope with citrate buffer, addition of blocking buffer for blocking, and addition of primary antibody VEGF (Bioss, BS1665R, 1 : 1000), CD34 (Bioss, BS5085R, 1 : 1000), and EGF (Bioss, BS1007R, 1 : 1000) and a secondary antibody (horseradish peroxidase-labeled goat rabbit IgG [H + L]; Beyotime, A0208). Finally, DAB substrate solution was applied to reveal the staining. Images were acquired using a Motic BA400 microscope (Motic, China).