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. 2015 Dec;53(6):834–843. doi: 10.1165/rcmb.2014-0376OC

Figure 1.

Figure 1.

Prostacyclin causes Ras-related protein 1 (Rap1)-dependent recruitment of krev interaction trapped-1 (KRIT1) gene to the cell cortical layer, localization to adherens junctions, and association with vascular endothelial (VE)-cadherin. (A) Human pulmonary endothelial cell monolayers grown on glass coverslips were transfected with nonspecific (top panels) or Rap1-specific small interfering RNA (siRNA) (bottom panels) for 48 hours and treated with vehicle or prostacyclin (PC) (200 ng/ml, 15 min) followed by immunofluorescence staining for KRIT1. Scale bar = 10 μm. (B) Translocation of KRIT1 to the membrane/cytoskeletal fraction was detected with specific antibodies. (C) Coimmunoprecipitation assay using antibody to VE-cadherin was performed, and VE-cadherin and KRIT1 content in the immunoprecipitates was detected using appropriate antibody. siRNA-induced Rap1 knockdown was monitored by Western blot with Rap1 antibody. Results are representative of three independent experiments. CTSK, cytoskeletal; IP, immunoprecipitation; nsRNA, negative strand RNA; si-Rap1, Rap1-specific siRNA; VEC, VE-cadherin; Veh, vehicle.