Fig. 9.

Analysis of steady-state levels of mRNA for taurine transporter (TAUT) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in control (−SIN-1) and SIN-1 treated (+SIN-1) ARPE-19 cells. Confluent cells were treated with or without 1 mM SIN-1 for 24 h. Poly(A)⁺ RNA was then isolated from these cells and used for semiquantitative RT-PCR. Primer pairs specific for human TAUT mRNA and human GAPDH mRNA were used. RT-PCR was done with a wide range of PCR cycles (9–30). Resultant products were run on a gel and then subjected to Southern hybridization with ³²P-labeled cDNA probes specific for TAUT and GAPDH. Hybridization signals were quantified using the STORM PhosphorImaging System, and intensities that were in the linear range with the PCR cycle number were used for analysis. GAPDH-specific band intensity was used as an internal control. Ratios of TAUT-specific band intensity to GAPDH-specific band intensity were then compared between control and SIN-1 treated cells. Ratio in control cells was taken as 1.