Fig. 2.

Endogenous TRPP1 and the cytoplasmic COOH terminus of PC1 (PC1ctt) colocalize and interact in the plasma membrane of renal vascular smooth muscle cells (RVSMCs) of neonatal pigs. A: Immunofluorescence staining of a RVSMC, demonstrating that both TRPP1 and PC1ctt are expressed in the plasma membrane and colocalize. B and C: In situ proximity ligation assay (PLA) detected an interaction between PC1ctt and TRPP1 in RVMSCs. Whereas PLA signals (red fluorescence) were absent in negative control cells labeled only with anti-TRPP1, the signals were detected in cells labeled with anti-PC1ctt and anti-TRPP1 antibodies. PLA was performed using the Duolink in situ PLA kit (Olink Bioscience, Uppsala, Sweden), as we have previously described (Adebiyi, 2014). Images were acquired with a Zeiss LSM Pascal laser-scanning confocal microscope. The rabbit anti-PC1ctt antibody was a kind gift from Dr. Oxana Beskrovnaya (Chauvet et al., 2004) (Genzyme Corporation). Bar = 10 μm.