一、 轉(zhuǎn)染小鼠胚胎成纖維細(xì)胞（NIH-3T3），轉(zhuǎn)染siRNA文章概述：

當(dāng)NIH/3T3細(xì)胞生長密度達(dá)到60%，在不含F(xiàn)BS培養(yǎng)基的六孔板中進(jìn)行轉(zhuǎn)染。對于細(xì)胞轉(zhuǎn)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(yīng)對照，使用 Advanced DNA/RNA轉(zhuǎn)染試劑(#AD600025 Zeta life, USA) 對細(xì)胞轉(zhuǎn)染；在轉(zhuǎn)染24小時(shí)后分別對TRIM11或ALKBH5通過蛋白質(zhì)印跡或qPCR檢測敲低效率。

二、其它

1、NIH/3T3細(xì)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(xué)公共衛(wèi)生學(xué)院毒理學(xué)教研室、河北醫(yī)科大學(xué)環(huán)境與健康重點(diǎn)實(shí)驗(yàn)室

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

三、轉(zhuǎn)染NIH-3T3細(xì)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(xué)舊金山校區(qū)聯(lián)合開發(fā)用于哺乳動物細(xì)胞、活體動物轉(zhuǎn)染的 Advanced DNA RNA 第三代多肽小分子轉(zhuǎn)染試劑，此技術(shù)成為新的蛋白功能、免疫細(xì)胞及干細(xì)胞治療、研發(fā)及生產(chǎn)的主要關(guān)鍵技術(shù)之一。

五、產(chǎn)品信息