title: RNA-seq of adult cardiac fibroblasts (aCFs), aCFs treated with TNF-α and IL-4 (aCF+CKs), and aCF+CKs expressing CD90 and VCAM1 (aVCFs)
design: RNA-seq of adult cardiac fibroblasts (aCFs), aCFs treated with TNF-α and IL-4 (aCF+CKs), and aCF+CKs expressing CD90 and VCAM1 (aVCFs)
library_construction_protocol: Human adult cardiac fibroblasts (aCFs) were acquired from a commercial supplier; aCF+CKs were produced by addition of TNF-α and IL-4 into the culture medium; aVCFs were isolated from aCF+CKs by MACS against CD90 and VCAM1. Human cardiac fibroblasts were cultured in HFDM-1(+) medium supplemented with 1% (v/v) Newborn Calf Serum (NBCS). After initial expansion, aCFs were cultured with TNF-α and IL-4 in the medium. aCF-CKs were incubated with a biotin-conjugated anti-CD106 (VCAM1) or anti-CD90 (THY1) antibodies and anti-biotin microbeads for isolation of aVCFs by MACS. Total RNA was extracted using the RNeasy Plus Mini Kit following the manufacturer's instructions (Qiagen). The library was constructed using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs) by GENEWIZ.