Article Title: Topiramate Treatment Protects Blood-Brain Barrier Pericytes from Hyperglycemia-Induced Oxidative Damage in Diabetic Mice
Figure Lengend Snippet: Mitochondrial CA (mCA) in the primary cultured PC derived from mouse brain microvessels. A, Cerebral PC stained for α-SMA are green , nuclei stained with 4′,6-diamidino-2-phenylindole (DAPI) are blue . PC (1 × 10 4 ) were fixed on chamber slides with 4% paraformaldehyde in PBS, permeabilized, blocked with 0.3% Triton, 10% normal donkey serum (NDS) in PBS, and incubated with mouse antihuman α-SMA antibody at 10 μg/ml in 1% NDS in PBS. Secondary antibody was goat antimouse fluorescein isothiocyanate-conjugated IgG (1:200). After incubation and washing, slides were counterstained with DAPI (10 μg/ml) in mounting media. B, Transcripts of CA VA and VB in the PC and the brain. RNA isolated from PC was reversed transcribed and PCR amplified by mitochondrial CA VA-specific ( top ) and CA VB-specific ( bottom ) primers. Mouse brain RNA was used as a positive control. C, Mitochondrial CA VA and VB polypeptides in the PC and the brain. Proteins (50 μg) isolated from the PC were separated on polyacrylamide gels and probed with antimouse CA VA or VB antibody at 1:3000 dilution. Secondary antibody was goat antirabbit horseradish peroxidase (1:5000)-conjugated IgG. Proteins from the mouse brain were used as a positive control.
Article Snippet: The antihuman FVIII antibody (MAB3440) and goat antimouse IgG conjugated with fluorescein isothiocyanate or Rhodamine Red were from Millipore (Billerica, MA).
Techniques: Cell Culture, Derivative Assay, Staining, Incubation, Isolation, Polymerase Chain Reaction, Amplification, Positive Control