Fig. 3. Overexpression of TRPC1 protects SH-SY5Y neuronal cells from MPP⁺.

A, Western blot on crude membranes prepared from control SH-SY5Y cells, or TRPC1-overexpressing cells treated with MPP⁺ (250 μm) for 10 h. The upper blot was probed using anti-TRPC1 antibody, and the lower portion of the same blot was probed with anti-SERCA2B antibodies. IB, immunoblot. B, MTT assays performed on control and TRPC1-overexpressing cells treated with 250 μm of MPP⁺ treated for 10 h. C and D, Western blot on whole cell lysates prepared from control SH-SY5Y cells or TRPC1-overexpressing cells treated with MPP⁺ (250 μm) for 10 h as well as with TRPC1 down-regulated cells treated with MPP⁺. The lower panel in D shows the same blot reprobed with actin antibodies. The blot was probed using anti-α synuclein antibodies overnight followed by the addition of peroxidase-conjugated anti-goat secondary antibody. The proteins were detected using ECL and exposed to an x-ray film or Lumiimager.