Figure 5.

A, RT-PCR analysis of TRPC mRNA. Equal amounts of cDNA, prepared from total RNA of control and JOE OPCs, were added for each reaction. TRPC isoform-specific PCR primers (Table 1) and the semiquantitative RT-PCR conditions used for these experiments are described in Materials and Methods. β-Tubulin was used as internal standard, and data are representative of three independent experiments. B, Pure OPCs from control and JOE mice were transiently transfected with a combination of three different siRNA duplexes specific for TRPC1 (siTRPC1) and grown for 2 d in defined culture media. Semiquantitative RT-PCR analysis of TRPC1 mRNA expression in OPCs was performed using β-tubulin as internal standard. Data from three independent experiments are summarized based on the relative spot intensities and plotted as percentage of controls. C, OPCs were grown under conditions described above. For each lane, 50 μg of total protein extract was applied, and Western blots were performed as described in Materials and Methods. A representative Western blot for TRPC1 is shown, and data from three independent experiments are summarized based on the relative spot intensities and plotted as percentage of controls.