Mouse anti hif2α

The fresh kidney tumors of mice were fixed in 4% para-formaldehyde and embedded in paraffin, then sectioned at a thickness of 5 μm. After dewaxing and rehydration, sections were treated with 10% H₂O_2. After 5% BSA blocking, all sections were incubated with the primary antibodies at 4°C overnight. The primary antibody was recognized by the biotinylated secondary antibody (Vector), and visualized by VECTASTAIN ABC peroxidase system and peroxidase substrate DAB kit (Vector). The primary antibodies used were: rabbit anti-AR (Santa Cruz), rabbit anti-MMP9 (Abcam), mouse anti-HIF2α (Abcam), rabbit anti-VEGF (Abcam) and rabbit anti-CCND1 (Cell Signaling).

Dissected livers were fixed in 4% paraformaldehyde in PBS at 4°C overnight and processed for paraffin embedding in a Leica ASP200S tissue processor. Histological analyses and Sirius red staining were performed as described previously (10 (link)).
The following primary antibodies were used at the indicated dilutions: rabbit anti–cleaved caspase-3 (1:200, Cell Signaling Technology, 9661); rat anti-CD45 (1:200, BD Pharmingen, BD Biosciences 557390); rabbit anti–collagen IV (1:400, Abcam, Ab19808); goat anti-GFP (1:200, Abcam, Ab6673); mouse anti-GATA4 (1:100, Santa Cruz Biotechnology, SC-25310); goat anti-HNF4α (1:100, Santa Cruz Biotechnology, SC-6556); rabbit anti-laminin (1:50, MilliporeSigma, L9393); mouse anti-HIF2α (1:100, Abcam, Ab8365); rabbit anti-Phospho-Histone H3 (1:500, MilliporeSigma, 06-570); mouse anti-Ki67 (1:100, Thermo Fisher Scientific, RM-9061); mouse anti–α-SMA (1:300, MilliporeSigma, A5228); rabbit anti-desmin (1:100, Abcam, Ab15200); band iotinylated lectin from Bandeiraea simplicifolia (L-3759, 1:100, MilliporeSigma). For image quantification, 20 random images at high magnification (40×) of 2 nonconsecutive sections of at least 3 mice per group were quantified using ImageJ software (NIH).