(a) Phenotypic susceptibility assay in presence of EtBr
to assess genotoxicity in C.
The percentage of EtBr
positive cells was significantly reduced by DPI or NAC treatment but the percentage of EthD-2 positive showed no significant difference (Figures 3(c)-3(d)).
To assess the potential activity of the 13 compounds as efflux pump inhibitors, a real-time EtBr
accumulation assay in a S.
DNA was stained with aqueous solution of EtBr
, visualized and photographed under a UV transilluminator.
The PCR products were analyzed by electrophoresis on 1.0% agarose gel and stained with ethidium bromide (EtBr
) and were detected by Gel imaging system.
The gels were stained in EtBr
and visualized with alpha imager gel documentation system.
For visualizing PCR products, 5 [micro]L of the amplified product was electrophoresed on 1% agarose gel in 1X TAE buffer, stained with ethidium bromide (EtBr
0.5 [micro]g/mL) and analyzed by gel documentation system (BIORAD).
After the run, gel was removed and stained with 10 [micro]g/mL ethidium bromide (ETBR
) for 10-15 min and the image was taken in UV transilluminator and photographed to determine the extent of DNA cleavage.
To understand the mechanism for the effect of rosiglitazone on ET-1-induced vasocontraction in pulmonary arteries, we examined the protein level of ETbR
with Western blotting.
The integrity of total RNA was assessed by running an aliquot of the RNA sample on a denaturing agarose gel stained with ethidium bromide (EtBr
) and by spectrophotometric analysis (Ultraspec 2100 pro Amersham Biosciences, USA).
Agarose gel for Electrophoresis: Weighed 0.8g of agarose in 40ml of 1X TBE Buffer in a micro-oven and added 4 [micro]L of Ethidium Bromide (EtBr
) as soon as it is removed from oven and mixed to make it uniform.