GSM6788494: T45; Homo sapiens; miRNA-Seq
For IPF fibrotic foci, Laser Capture Microdissection was used to isolate fibrotic foci from formalin embedded IPF tissue. RNA was isolated from LCM cut tissue and whole IPF formalin embedded tissue using FFPE RNeasy extraction kit (Qiagen). Lung fibroblasts were isolated from IPF lung tissue and normal human lungs. The cells were grown to 90% confluence, then serum starved for 24h in media containing 0.4% foetal calf serum. After the 24h perios, the cells were incubated for 48h with complete media only or media supplemented with 3ng/ml TGFβ1. The cells were harvested at 48h and total RNA extracted using an RNeasy mini kit (Qiagen). NEBNext small RNA libraries (New England BioLabs Inc.) were prepared from total RNA.