图4. 邻近标记中生物素浓度与处理时间的优化
(A) 使用不同浓度的生物素溶液分别对野生型和转TurboID-EYFP拟南芥处理1小时, 使用SA-HRP和anti-GFP抗体进行免疫印迹检测, 分析TurboID-EYFP的活性与表达水平; (B) 用50 µmol∙L^-1生物素溶液处理野生型和转TurboID-EYFP拟南芥不同时间, 然后抽提蛋白并进行免疫印迹分析。SA-HRP: 辣根过氧化物酶标记链霉亲和素; CBB: 考马斯亮蓝

Figure 4. The optimization of biotin concentration and labeling time used in proximity labeling
(A) Wild-type (WT) and transgenic Arabidopsis seedlings overexpressing TurboID-EYFP were treated with different concentrations of biotin solution for 1 h, followed by immunoblots with SA-HRP and anti-GFP antibodies to detect the proximity ligation activity and protein expression level of TurbolD-EYFP; (B) Wild-type (WT) and transgenic Arabidopsis seedings overexpressing TurboID-EYFP were treated with 50 µmol∙L^-1 biotin for different times, followed by immunoblots with appropriate antibodies. SA-HRP: Streptavidin-HRP; CBB: Coomassie Brilliant Blue