Author: Li, Zi; Lan, Yungang; Zhao, Kui; Lv, Xiaoling; Ding, Ning; Lu, Huijun; Zhang, Jing; Yue, Huiqing; Shi, Junchao; Song, Deguang; Gao, Feng; He, Wenqi
Title: miR-142-5p Disrupts Neuronal Morphogenesis Underlying Porcine Hemagglutinating Encephalomyelitis Virus Infection by Targeting Ulk1 Document date: 2017_5_3
ID: 07b3pbxc_13
Snippet: Primary cortical neuron lysate or freshly isolated cerebral cortex lysate was diluted in RIPA lysis buffer on ice. Subsequently, SDS-PAGE was performed, and the lysate samples were electrotransferred to 0.22 µm polyvinylidene difluoride membranes using the Bio-Rad wet transfer system. And then, the membranes were blocked overnight at 4 • C with 5% non-fat dry milk in PBS and immunoblotted with antibodies against Ulk1 (Cell Signaling Technologi.....
Document: Primary cortical neuron lysate or freshly isolated cerebral cortex lysate was diluted in RIPA lysis buffer on ice. Subsequently, SDS-PAGE was performed, and the lysate samples were electrotransferred to 0.22 µm polyvinylidene difluoride membranes using the Bio-Rad wet transfer system. And then, the membranes were blocked overnight at 4 • C with 5% non-fat dry milk in PBS and immunoblotted with antibodies against Ulk1 (Cell Signaling Technologies, USA, 1:1,000), β-actin (Proteintech, USA, 1:2,000) and PHEV (provided by our laboratory, 1:500) at 4 • C overnight. Next, incubation with horseradish peroxidase (HRP)-conjugated secondary anti-rabbit or anti-mouse IgG antibodies (Proteintech, USA) was performed for 1 h at 37 • C.
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