《Nanoparticles in Autophagy Modulation》

• 来源专题：现代化工
• 编译者： 武春亮
• 发布时间：2024-06-19
• 原文来源：https://www.mdpi.com/2079-4991/14/12/1033

LC3 transfection was conducted using GFP–LC3 Expression Vector (CELL BIOLABS, San Diego, CA, USA, CBA-401). Briefly, experiments were performed after growing at 80–90% cell confluence. Cells were detached by Trypsin-EDTA (0.25%) (GibcoTM, 25200072) and centrifuged at 1200 rpm for 3 min. The supernatant was discarded and resuspended in new media. The cells were collected (0.5 × 10
6
cells) and centrifuged (300 g for 5 min). The supernatant was discarded and washed with DPBS (WELGENE, Busan, Republic of Korea, LB 001-02). After centrifuge, cells were resuspended with DPBS. Then, 1 μg GFP-LC3 DNA was added to the cells. Transfection was performed with the NeonTM Transfection System, which is an electroporation system (InvitrogenTM, Carlsbad, CA, USA). After transfection, the cells were seeded to a 6-well plate without antibiotic. Selection cells were proceeded with GeneticinTM Selective Antibiotic (G418 Sulfate; GibcoTM, 10131035). G418 was treated at 800 μg/mL for 3 days and changed to a decreased concentration (200 μg/mL).