Transfection and Gene Silencing in Pericytes

Pericytes were transfected using Amaxa Nucleofector Technology and the Basic Nucleofector Kit (Lonza, Switzerland, EU, Cat# VPI-1001) as described [15 (link)]. For ocln overexpression, cells were transfected with 1 μg PCMV3-OCLN plasmid (Sino Biological, Wayne, PA, Cat# HG15134-UT) or with PCMV3 (Sino Biological, Cat# CV011) as a negative control. For gene silencing, cells were transfected with 0.5 μg per 10⁶ cells of the following small interfering RNA (siRNA): 3 unique 27mer OAS1-siRNAs (OriGene, Rockville, MD, USA, Cat# SR303266), 3 unique 27mer OAS2-siRNAs (OriGene, Cat# SR303267), 3 unique 27mer OAS3-siRNAs (OriGene, Cat# SR303268), 3 unique 27mer OASL-siRNAs (OriGene, Cat# SR305676), 3 unique 27mer RNaseL-siRNAs (OriGene, Cat# SR304081), 3 unique 27mer occludin-siRNAs (OriGene, Cat# SR303274), 3 unique 27mer TJP1-siRNAs (OriGene, Cat# SR322042), or trilencer-27 universal scrambled (SCR) silencer siRNA duplex (OriGene, Cat# SR30004) as a negative control.