Cardiac NRVM Culture and RNA-seq Analysis

Cardiac NRVMs were grown on glass or 10 kPa substrates with fibronectin or fibronectin and TTR fibrils for 72 h. Total RNA was purified using Maxwell^® RSC simplyRNA Cells (Promega) with the inclusion of a DNAse treatment step. RNA samples were quantified using NanoDrop™ One Spectrophotometer (Thermo Scientific) and analyzed for integrity using Agilent 4200 TapeStation. Levels of remaining DNA were checked using a Qubit fluorometer (Invitrogen). Sequencing and gene expression statistical analysis were performed as previously described (Dittloff et al., 2021 (link)). Pathway analysis of differentially expressed genes was performed in Ingenuity Pathway Analysis. Visualization of differential expression and pathway analysis was performed via Integrated Differential Expression and Pathway analysis (Ge et al., 2018 (link)). Data are available in Figure S1–S3.