GSM5207234: E3_015Chip8_FPKM; Homo sapiens; RNA-Seq
Laser capture microdissection was performed upon formalin fixed paraffin embedded (FFPE) control non-fibrotic lung tissue (alveolar septae (n=10)) and UIP/IPF FFPE lung tissue (myofibroblast foci, (n=10) and adjacent non-affected alveolar septae (n=10)). Regions of interest (non-fibrotic control lung tissue alveolar septae, IPF lung tissue fibroblast foci or adjacent non-affected alveolar septae) were selected using a drawing tool on the Leica Laser Microdissection software, and then cut with a laser beam into separate RNase-free tubes containing RecoverAll Total Nucleic Acid digestion buffer (Ambion). RNA isolation was performed using Micro Filter Cartridges from RNAqueous-Micro Total RNA Isolation Kit (Invitrogen) according to the manufacturer's instructions. 15ng of total RNA was used to make cDNA libraries using Ion Ampli‐Seq‐transcriptome human gene expression kit (Ion Torrent, Life Technologies). An Ion Torrent Proton Sequencer (RNA-Seq) was used to produce ~20 million single-end ~100bp reads/sample.