Expression profile of knockout Mettl3 in CD4+ T cells with Imiquimod induced psoriasis mouse model

In the psoriasis mouse model, depletion of the RNA N6-methylase, Mettl3, from CD4+ T cells inhibits psoriasis-like phenotype and inflammation. To isolate CD4+ T cells, mouse dorsal skin was excised and the fat layer was removed using a surgical scalpel. The skin tissue was treated with dispase II (2.5 mg/ml) in Hank’s Balanced Salt Solution for 60 min at 37 °C. The dermal fraction was collected by scraping and then cut into small pieces. Dermal single-cell suspensions were obtained after collagenase IV (1 mg/ml) treatment for 2 h at 37 °C, followed by filtering and centrifugation. Samples were stained for 30 min on ice. The following antibodies were used: anti-CD4-PE (100408, Biolegend) and anti-CD45-FITC (103108, Biolegend).

在银屑病小鼠模型中，从CD4+ T细胞中敲除RNA N6-甲基转移酶（RNA N6-methylase）Mettl3，可抑制银屑病样表型与炎症反应。为分离CD4+ T细胞，对小鼠背部皮肤进行取材并使用手术手术刀去除脂肪层。将皮肤组织置于添加有2.5 mg/ml分散酶II（dispase II）的汉克斯平衡盐溶液（Hank’s Balanced Salt Solution）中，于37℃孵育60分钟。通过刮取收集真皮组分，随后将其剪切成小块。使用浓度为1 mg/ml的IV型胶原酶（collagenase IV）于37℃孵育2小时，经过滤与离心后制备得到真皮单细胞悬液。样本于冰上染色30分钟，所用抗体包括：抗CD4-PE（货号100408，Biolegend）与抗CD45-FITC（货号103108，Biolegend）。