Illumina NovaSeq 6000 paired end sequencing; Influence of substrate stiffness on barrier function in an iPSC-derived in vitro blood-brain barrier model
Influence of substrate stiffness on barrier function in an iPSC-derived in vitro blood-brain barrier model
On day 7after purification, BMEC-like cells were detached from PA hydrogel surfaces with a 10 minuteAccutase incubation at 37°C. Cells from 3 gels of each stiffness condition were pooled to create one biological replicate. Cells were then rinsed with PBS and incubated in Trizol (Thermo Fisher 15596026) for 10 minutes at room temperature. RNA was extracted using a Direct-zol RNA miniprep kit (Zymo R2050) according to the manufacturer's instruction. This process was repeated two additional times from independent iPSC differentiations (biological n=3).  Raw sequencing reads were obtained for the 9paired-end samples. FASTQ reads were mapped to human genome (hg19) by HISAT2 (version 2.1.0)