GSM8162560: Mettl3-etoh-Nec-1-2; Mus musculus; RNA-Seq
organoids were harvested and collected by centrifugation. Total RNA was extracted with RNeasy Mini Kit (QIAGEN) according to the manufacturer's instructions. The RNA-seq library were prepared for sequecing using NEBNext Ultra RNA library Prep Kit for Illumina. For ChIP-seq library, Alive lentivirus-infected organoids overexpressing Atf3 were sorted and collected into 500 uL PBS, cells conducted ChIP-seq following the NovoNGS CUT&Tag 3.0 High-Sensitivity Kit (Novoprotein, H259-YH01) and the lysates were incubated with anti-ATF3 antibody (1:100, Abcam, ab207434) at 4 ℃ overnight. DNA extracted using Tagment DNA Extract Beads was used as a templet, and amplification was performed with 2X KAPA HIFI HotStart polymerase (KAPA, KK2601).The library was purified with 1.4X AMPure beads and sequenced with the Illumina Novaseq 6000.