Single cell RNA-sequencing of inguinal adipose tissue Pdgfrβ+ cells

We previously derived a doxycycline-inducible (Tet-On) lineage-tracing model that allows for the indelible labeling of Pdgfrb-expressing perivascular cells in adipose tissue of adult mice (PdgfrbrtTA; TRE-Cre; Rosa26RmT/mG; herein, MuralChaser mice). Prior to exposing animals to doxycyline, all cells within the stromal-vascular fraction (SVF) of adult inguinal WAT (iWAT) express membrane tdTomato from the Rosa26 locus. Following 9 days of exposure to doxycycline-containing chow diet, Cre-mediated excision of the loxP-flanked tdTomato cassette occurs in Pdgfrb-expressing cells, and membrane-bound GFP (mGFP) expression is constitutively activated. We set out to test the hypothesis that Pdgfrb-expressing perivascular cells in inguinal WAT of adult mice are heterogeneous, with subpopulations harboring functionally distinct phenotypes. To this end, we performed single cell RNA-sequencing of mGFP+ cells isolated from iWAT of lean (chow fed) 8 weeks-old male MuralChaser mice following 9 days of doxycycline exposure. We pooled tdTomato- GFP+ cells from inguinal white adipose tissue (WAT) from 5 MuralChaser mice. Single cell library preparation was performed using the 10X Genomics Single Cell 3 v2 according to the manufacturers instructions.