一、 轉染小鼠胚胎成纖維細胞（NIH-3T3），轉染siRNA文章概述：

當NIH/3T3細胞生長密度達到60%，在不含F(xiàn)BS培養(yǎng)基的六孔板中進行轉染。對于細胞轉染使用5μM pGPU6/GFP/Neo-shTRIM11,siALKBH5,miR-21a-5p mimic,miR-590–5p mimic, miR-361–3p mimic, miR-202–3p mimic和相應對照，使用 Advanced DNA/RNA轉染試劑(#AD600025 Zeta life, USA) 對細胞轉染；在轉染24小時后分別對TRIM11或ALKBH5通過蛋白質印跡或qPCR檢測敲低效率。

二、其它

1、NIH/3T3細胞培養(yǎng)條件：文章選擇Z7181FBS-500胎牛血清培養(yǎng)(# Zeta life, USA)

2、文章作者：河北醫(yī)科大學公共衛(wèi)生學院毒理學教研室、河北醫(yī)科大學環(huán)境與健康重點實驗室

3、標題：FcγRIIB The proteasome-dependent degradation of ALKBH5 regulates ECM deposition in PM2.5 exposure-induced pulmonary fibrosis of mice .(IF10.588)

三、轉染轉染NIH-3T3細胞原文

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.

四、美國Zeta Life 公司與美國加利福尼亞大學舊金山校區(qū)聯(lián)合開發(fā)用于哺乳動物細胞、活體動物轉染的 Advanced DNA RNA 第三代多肽小分子轉染試劑，此技術成為新的蛋白功能、免疫細胞及干細胞治療、研發(fā)及生產(chǎn)的主要關鍵技術之一。

五、產(chǎn)品信息