The m6A methyltransferase Mettl3 deficiency attenuates hepatic stellate cell activation and liver fibrosis [RNA-seq]

RNA sequencing of LX2 cell line (a human hepatic stellate cell line). METTL3 overexpression or downreguation were achieved by lenti-virus transduction. Two shRNAs targeting human METTL3 (shMETTL3-1 and shMETTL3-2) or firefly luciferase (shLuc) were cloned into pLKO.1 lentiviral vector (Addgene, 10878). shMETTL3 was used to achieve METTL3 knockdown, and shLuc was used as control. Lentivirus transfections were performed using polyethylenimine (PEI, Polysciences, 23966) according to the manufacturer's protocol. LX2 was infected with lentivirus-containing supernatants. Successfully infected cells were selected and prepared for futher RNA-seq.

本数据集针对人肝星状细胞系LX2开展RNA测序（RNA sequencing）。通过慢病毒转导（lenti-virus transduction）实现METTL3的过表达与敲低。将两条靶向人METTL3的短发夹RNA（short hairpin RNA，shRNA，分别为shMETTL3-1与shMETTL3-2），以及靶向萤火虫荧光素酶的shLuc，均克隆至pLKO.1慢病毒载体（Addgene，10878）中。其中shMETTL3用于介导METTL3基因敲低，shLuc则作为阴性对照。采用聚乙烯亚胺（polyethylenimine, PEI，Polysciences，货号23966），并依照制造商提供的实验规程进行慢病毒包装转染。将LX2细胞与含重组慢病毒的细胞上清共孵育以完成感染，筛选获得成功感染的细胞并制备样本，用于后续RNA测序（RNA-seq）。