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_15
Snippet: Primary cortical neurons were fixed in 4% paraformaldehyde (Fisher Scientific, Waltham, MA), permeabilized with 0.2% Triton X-100 and blocked in bovine serum albumin (BSA). Immunostaining was performed overnight with microtubuleassociated protein 2 (MAP2) (D5G1) XP R rabbit mAb (Cell Signaling Technologies, USA), ULK1 (D8H5) rabbit mAb, and/or PHEV mouse mAb (provided by our laboratory) in 1% BSA at 4 • C. Alexa Fluor 568-or Alexa Fluor 488-con.....
Document: Primary cortical neurons were fixed in 4% paraformaldehyde (Fisher Scientific, Waltham, MA), permeabilized with 0.2% Triton X-100 and blocked in bovine serum albumin (BSA). Immunostaining was performed overnight with microtubuleassociated protein 2 (MAP2) (D5G1) XP R rabbit mAb (Cell Signaling Technologies, USA), ULK1 (D8H5) rabbit mAb, and/or PHEV mouse mAb (provided by our laboratory) in 1% BSA at 4 • C. Alexa Fluor 568-or Alexa Fluor 488-conjugated antimouse or anti-rabbit secondary antibodies (Invitrogen, 1:1,000) were applied for 1 h at room temperature. After counterstaining with 1 µg/mL Hoechst 33342 (Sigma) for 2 min, coverslips were mounted with ProLong Gold Antifade Reagent (Life Technologies), and the slides were observed under a confocal microscope (FV10-ASW 3.0, Olympus Europa Holding GmbH).
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