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Fig. 2 | BMC Obesity

Fig. 2

From: Adipocyte nuclei captured from VAT and SAT

Fig. 2

Expression of reporters and capture SUN1mRFP1Flag tagged nuclei from mature 3T3-L1 adipocytes. a Six days after ADNp::mRFP1 nucleofected 3T3-L1 preadipocytes were induced to differentiate into MAs strong cytoplasmic fluorescence of the mRFP1 reporter was detected (see Table 1, Additional file 2: Figure S2). Pre-adipocytes did not have detectable fluorescence. This result shows the promoter vector was specifically expressed in MAs. b mRFP1 fluorescence from the ADNp::SUN1mRFP1Flag reporter on the surface of differentiated 3T3-L1 nuclei selected from c. c Expression of the ADNp::SUN1mRFP1Flag reporter in MAs 10 days after induction of nucleofected 3T3-L1 preadipocytes (DAPI stained DNA (blue), lipid rich oil droplets detected with Lipotox Green (green), enhanced red fluorescent protein mRFP1 (red). The lower right hand panel shows the merged fluorescence image. mRFP1 fluorescence is associated with the nuclei. It should be noted that spherical oil droplets often produce an optical distortion of adjacent nuclei. d Adipocyte nuclei from mature 3T3-L1 cells expressing the SUN1mRFP1Flag reporter were captured with Protein-G Dynabeads (2.8 μm diameter) pre-adsorbed with anti-GFP antibody. e A negative control capture performed in parallel using Protein-G Dynabeads pre-adsorbed with anti-CD4 T cell specific antibody did not capture nuclei

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