TRPC6 in the neurons was downregulated in ischemia.
(A) Immunoblots of the extracts from the contralateral (C) or ipsilateral (I) cortex (sham; left [L] or right [R] hemisphere) using the indicated antibodies. Tubulin served as a loading control. Right panel: quantification of the normalized TRPC6 protein levels (n = 5–8 rats per time point). *P < 0.05; **P < 0.01 versus sham. (B) Immunoblots for TRPCs and GluR1 after 24 hours reperfusion (R24). Right panel: quantification of the protein levels (n = 5 rats). **P < 0.01 versus contralateral. (C) TRPC6 mRNA levels determined by qRT-PCR (n = 3–5 rats per time point). (D) Representative images of the indicated cortex after 24 hours reperfusion (R24) double-stained with the indicated antibodies. Scale bar: 50 μm. Quantification of the optical density for both TRPC6 and NeuN immunoreactivity is shown on the right panel (n = 5 rats). **P < 0.01 versus the density in contralateral cortex. Data are presented as mean ± SEM. Unless stated, TRPC6 antibody from Millipore was used for Figures Figures1,1, ,2,2, and and66 and antibody from Alomone Labs was used for Figures Figures3,3, ,4,4, ,5,5, and and7.