Immunostaining was performed as previously described in detail (Jumabay et al., 2012 (link)). Briefly, cells grown in chamber slides were fixed in 4% paraformaldehyde, permeabilized with 0.2% Triton X-100, blocked with 10% goat serum and 1% BSA in PBS, and incubated over night at 4°C with the appropriate primary antibodies or non-specific IgG control antibodies, diluted 1:200 in 1% BSA in PBS. The next day, cells were incubated with secondary AF-488-conjugated (green fluorescence) or AF-594-conjugated (red fluorescence) goat anti-mouse or anti-rabbit secondary antibodies (Molecular Probes). The cells were washed with PBS, the nuclei stained with 4',6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich), and visualized by fluorescence microscopy. The non-specific IgG control antibodies showed no staining and are not included in the figures.
We used the following antibodies for immunostaining: hamster anti-CD31, rabbit anti-vone Willebrand Factor (vWF) (both from Dako), goat anti-BMP4, goat anti-MGP, rabbit anti-VEGF, rabbit anti-VE-Cadherin (all from Santa Cruz Biotechnology), mouse anti-Perilipin (Cell Signaling Technology).
We used the following antibodies for immunostaining: hamster anti-CD31, rabbit anti-vone Willebrand Factor (vWF) (both from Dako), goat anti-BMP4, goat anti-MGP, rabbit anti-VEGF, rabbit anti-VE-Cadherin (all from Santa Cruz Biotechnology), mouse anti-Perilipin (Cell Signaling Technology).
