GSM9181658: 8025, Arpc4 KO-3, 75% Matrigel; Mus musculus; RNA-Seq
Total RNA was extracted by adding TRIzol reagent (Thermo Fisher, USA) to the gel after removing the supernatant and dissolving the gel by thorough mixing. Chloroform (Thermo Fisher, USA) was then added, followed by vigorous vortexing for 15 seconds. After incubation at room temperature for 3 minutes, the mixture was centrifuged at 12,000×g for 15 minutes at 4°C. The upper aqueous phase was carefully transferred to a fresh tube, mixed with an equal volume of ≥95% ethanol, and purified using the Monarch Total RNA Miniprep Kit (New England Biolabs, Germany) according to the manufacturer's protocol. RNA sequencing was performed by Biomarker Technologies(BMKGENE), Münster, Germany. The concentration of extracted nucleic acid was detected with Nanodrop2000, , and the integrity was detected with Agient2100, LabChip GXPerkin Elmer LabChip GX). (manufacturer and model:Perkin Elmer LabChipGX). VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina NR605 was used to construct the mRNA library and strand-specific mRNA library in strict accordance with the protocol provided by Vazyme. The library preparations were sequenced on the Illumina Novaseq X platform (Illumina, SanDiego, CA) and reads were generated.