title: GSM8689179: CRS_6; Mus musculus; RNA-Seq
library_construction_protocol: Chronic rhinosinusitis (CRS) mice model To establish CRS mice model, mice in CRS group were treated intranasally with 20μg Papain (Sigma-Aldrich, P5306-25MG, USA) which was dissolved in 20μL phosphate buffer saline (PBS) on days 0-2 and 7-11. Drug treatment The treatment dosage of C10 (MCE, HY-19357, CHN) drug solution for intranasal administration in CRS mice were 10 mg/mL, with bilateral administration (10 μl/side, bilateral intranasal instillation). Starting from the initial administration of papain, treatment was continued for 10 consecutive days, with once-daily application. The concentration 10 ng/mL of IL-13 (MCE, HY-P72795, CHN) was used to stimulate Beas-2b and HNECs for 48 hours. A total amount of 1-3μg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using VAHTS Universal V6 RNA-seq Library Prep Kit for Illumina ® （NR604-01/02）following the manufacturer's recommendations and index codes were added to attribute sequences to each sample. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Then we added fragmentation buffer to break the mRNA into short fragments. First strand cDNA was synthesized using random hexamer primer and RNase H. Second strand cDNA synthesis was subsequently performed using buffer, dNTPs, DNA polymerase I and RNase H. And then, the double stranded cDNA was purified by AMPure P beads or QiaQuick PCR kit. The purified double stranded cDNA was repaired at the end, added a tail and connected to the sequencing connector, then the fragment size was selected, and finally the final cDNA library was obtained by PCR enrichment.