RNAseqofA549cell cultured in soft ECM
Bioprinted samples were first digested to collect the cells. 2D cultured cells were digested from culture flasks and centrifuged to collect a pellet. Trizol was immediately added to cell pellets to preserveRNAintegrity. Total RNA was extracted using Zymo Microprep kit. MessengerRNAwaspurifiedfromtotalRNAusingpoly-Toligo-attachedmagneticbeads.Afterfragmentation,thefirststrandcDNAwassynthesizedusingrandomhexamerprimersfollowedbythesecondstrandcDNAsynthesis.Thelibrarywasreadyafterendrepair,A-tailing,adapterligation,sizeselection,amplification,andpurification.