GSM5013354: Bulk RNA Seq of FACS sorted P3 Inguinal White Adipose Tissue (IWAT) ICAM1+ PreAdipocytes [ICAM11]; Mus musculus; RNA-Seq
The inguinal white adipose tissue and the aorta along with associated perivascular adipose tissue were isolated from P3 mice (Biological replicates represent tissues from the same pools of mice. I.e. PVAT_Rep1 and Inguinal_Rep1 contain tissue from the same individuals). Tissues were minced and placed into digestion medium (DMEM, Collagenase D: 6.1mg/ml (Roche), Dispase II: 2.4 mg/ml (Roche) and placed at 37C with constant agitation at 200 rpm. FACS was performed as decribed in the manuscript to isolate intermediate cells, progenitor cells, preadipocytes, and smooth muscle cells. Cells were sorted into FBS catch buffer. After sorting, cells were spun down and the cells were resuspended in 250uL of FACS buffer. Cells were then lysed with the addition of 750uL of Trizol LS (Invitrogen) and RNA was purified by phenol chloroform extraction. Genewiz Ultra Low Input Library Protocol "RNA was quantified using Qubit and TapeStation. RNA sequencing libraries were made using the SMART-Seq v4 Ultra Low Input Kit. This library prep uses Poly(A) selection to target mRNA, with enrichment for full length transcripts."