The immunocytochemical staining was performed as described earlier (Huttala et al., 2015) (link) except that the fixative used here was 4% formaldehyde at room temperature for 20 min. Antibodies used were anti-von Willebrand factor IgG (produced in rabbit, Sigma), anti-collagen IV IgG (produced in mouse, Sigma), FITC-labeled goat polyclonal anti-mouse IgG (Sigma), and TRITC-labeled goat polyclonal anti-rabbit IgG (Sigma). In addition, for confocal imaging, anti-collagen IV (produced in rabbit, ab6586, Abcam, Cambridge, UK) used with secondary antibody TRITC-labeled goat polyclonal anti-rabbit IgG (Sigma), CD140b-PE (BD Biosciences, 558821), and CD144-FITC (BD Biosciences, 560411) were used. Nuclei were stained using Flu-oroshield™ with DAPI mounting medium (Sigma).
Huttala O., Sarkanen J.R., Heinonen T, & Ylikomi T. (2019). Presence of vasculature results in faster insulin response in adipocytes in vascularized adipose tissue model. ALTEX, 36(3).
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