一、 转染小鼠胚胎成纤维细胞(NIH-3T3),转染siRNA文章概述:
当NIH/3T3细胞生长密度达到60%,在不含FBS培养基的六孔板中进行转染。对于细胞转染使用5μM pGPU6/GFP/Neo-shTRIM11,siALKBH5,miR-21a-5p mimic,miR-590–5p mimic, miR-361–3p mimic, miR-202–3p mimic和相应对照,使用 Advanced DNA/RNA转染试剂(#AD600025 Zeta life, USA) 对细胞转染;在转染24小时后分别对TRIM11或ALKBH5通过蛋白质印迹或qPCR检测敲低效率。
二、其它
1、NIH/3T3细胞培养条件:文章选择Z7181FBS-500胎牛血清培养(# Zeta life, USA)
2、文章作者:河北医科大学公共卫生学院毒理学教研室、河北医科大学环境与健康重点实验室
3、标题:FcγRIIB The proteasome-dependent degradation of ALKBH5 regulates ECM deposition in PM2.5 exposure-induced pulmonary fibrosis of mice .(IF10.588)
三、转染转染NIH-3T3细胞原文
Cell transfection
Transfection was conducted when NIH/3T3 cells grown to 60% density in six-well plates with FBS absent medium. For cell transfection, 5 μM of pGPU6/GFP/Neo-shTRIM11, siALKBH5, miR-21a-5p mimic, miR-590–5p mimic, miR-361–3p mimic, miR-202–3p mimic and cor-responding control (Genepharma, Shanghai, China) was transfected into cells using Advanced DNA/RNA Transfection Reagent (Zeta life, USA) according to the supplier’s guidelines, respectively. TRIM11 or ALKBH5 knockdown efficiency was detected by western blotting or qPCR after 24 h post-transfection, respectively.
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