Skip to main content
Fig. 1 | BMC Obesity

Fig. 1

From: Adipocyte nuclei captured from VAT and SAT

Fig. 1

Strategy for implementing INTACT to capture adipocyte nuclei. a The ADNp::SUN1mRFP1Flag gene construct. This promoter is a truncated 5.4 kb version of the mouse adiponectin promoter (ADNp) modified slightly from one described previously [63]. This promoter was fused first to the N-terminal portion of mouse SUN1, then via a flexible linker peptide to the enhanced red fluorescent protein mRFP1, which was fused in frame through a linker peptide to a epitope Flag tag containing three distinct Flag sequences (3xFlag). The ADIPOQ promoter region contains a 5´ upsteam exon (yellow) and intron, a 5´ mRNA UTR, and the ATG start codon. The 3´ UTR and poly(A) addition site are from SV40 in the pcDNA3.1 vector. More detailed maps are shown in Additional file 1: Figure S1 and Additional file 3: Figure S3. b Proposed deposition of the SUN1mRFP1Flag protein tethered to the inner nuclear membrane and tagging the exterior of the nucleus. c Diagram of adipose tissue comprised of MAs expressing the SUN1mRFP1Flag tag and many non-target cell types (progenitor cells, leukocytes, endothelial cells). d Capture and enrichment of MA nuclei away from other cellular nuclei on paramagnetic beads labeled with anti-mRFP1, anti-GFP or anti-Flag. A total of four constructs designed from 3 nuclear transmembrane proteins were examined (Table 1)

Back to article page