Anti uchl1

The recombinant rat TNF-α was purchased from R&D systems, Inc., Minneapolis, MN, USA. The Millicell was purchased from EMD Millipore Corporation, Billerica, MA, USA. The enhanced chemiluminescence detection kit and antibodies (anti-αsmooth muscle actin, anti-UCH-L1, anti-p65, anti-lamin a/ c, anti-GAPDH) were purchased from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA. The vectors of pDONR221 and pAd/CMV/V5-DEST, recombination system, BLOCK-iT Pol II miR RNAi Expression Vector Kit with EmGFP, and Lipofectamine 2000 were purchased from Invitrogen, Carlsbad, CA, USA. The ELISA kits for IL-6 and IL-8 were purchased from Thermo Scientific, Inc. Waltham, MA, USA. Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) were purchased from Hyclone, Cramlington, UK. Nuclear extract kit was purchased from Active Motif (Carlsbad, CA, USA). Cell culture: The primary rat VSMCs were isolated from thoracic aortic media of male Wistar rats (180-200 g) according to Sachinidis's method. 10) The cells were cultured in DMEM supplemented with 10% FBS, and maintained in a humidified atmosphere of 5% CO2 at 37℃. Following four passages, cells were immunostained with an anti-αactin antibody to characterize the cells as VSMCs by using the method of fluorescence immunoassay.