Fig. 3From: Adipocyte nuclei captured from VAT and SATMature MA-INTACT mouse adipocyte nuclei expressed SUN1mRFP1Flag RNA in BAT, VAT, and SAT allowing adipocyte nuclei to be captured on Immuno-paramagnetic beads. a Illustration of mouse fat deposits examined. b, c, d, e. Nuclei were efficiently captured from crude nuclear preparations prepared from inguinal SAT (b) retroperitioneal VAT (c) epididymal VAT (d) and scapular BAT (e). Protein G Dynabeads pre-adsorbed to rabbit anti-mRFP1 polyclonal antibody were used in these capture experiments. The clumping of nuclei occurs during successive rounds of washing and capture. Negative control antibodies did not capture significant numbers of nuclei (not shown). f A Western blot (top panel) showed that preparations of captured VAT, SAT, and BAT nuclei expressed the 131Â kDa SUN1mRFP1Flag reporter protein, while uncaptured nuclei did not. In house prepared anti-mRFP1 rabbit pAb detected the mRFP1 domain. Purified 25Â kDa mRFP1 was run as a positive control. A loading control (bottom panel) showed the approximately equal loading of nuclear proteins and an mRFP1 standard. The loading control samples were run for a short time (20Â min) on an SDS PAGE system and protein front stained with CoomassieBack to article page