Figure 1.

Immunostaining and western blotting to determine the location and expression, respectively, of CaSR and TRPC-1 in HUVECs. Confocal micrographs revealed that (A) CaSR protein was primarily expressed in the cytosol of HUVECs and (B) TRPC1 protein was expressed on the plasma membrane of HUVECs. Negative controls for (C) CaSR and (D) TRPC1 were performed using the secondary antibodies only. (E) Western blotting indicated that spermine treatment (2 mM, 48 h) did not alter the protein expression levels of CaSR and TRPC1 in HUVECs. Representative results from three independent experiments are presented. Magnification, ×200. Scale bar=50 µm. CaSR, Ca²⁺-sensing receptor; TRPC1, transient receptor potential canonical 1; HUVEC, human umbilical vein endothelial cells.