Immunohistochemistry and Immunofluorescence of AGGF1, VEGF, and CD34

Immunohistochemistry and immunofluorescence were performed as described in our previous study [19 (link), 20 (link)]. Primary antibodies were rabbit anti-AGGF1 (ab203680, 1:200), mouse anti-VEGF (ab1316, 1:150) and mouse anti-CD34 (ab8536, 1:400, all from Abcam, USA). The HRP-conjugated second antibody was from Invitrogen, Carlsbad, CA and the diaminobenzidine was from Beijing Zhongshan Golden Bridge Biotech, China. As for immunofluorescence, the primary antibodies cocktail and the mixture of secondary antibodies were rabbit anti-AGGF1 (1:150), mouse anti-CD34 (1:200) and Alexa Fluor 488-conjugated donkey anti-rabbit, Alexa Fluor 568 donkey anti-goat (all from Invitrogen), respectively.
For the negative control, the primary antibody was carried out with phosphate-buffered saline (PBS).