Intracellular Rap1 retention is prevented by TRPM8 silencing in ECs. (A) Live-cell imaging by confocal microscopy of ECs expressing GFP-Rap1 (green) and ER-DsRed (red) in control (siCNTRL) and siTRPM8-treated ECs (siTRPM8). Representative images showing Rap and ER dynamics in siCNTRL- and siTRPM8-treated ECs at t = 0 and after 2 or 10 min of treatment with 10 µM icilin; dotted white lines represent the PM. Representative plot profiles along the white lines in the images. The green bars depict the distance between the Rap1 and ER fluorescence signals. (B) Plot showing the means from a representative experiment of the distances measured between Rap1 and ER between Rap1 and TRPM8 in control (siCNTRL) and siTRPM8-treated ECs treatment with 10 µM icilin. The distances were calculated as means of at least three different plot profiles for at least three ROIs (cross sections) within each EC.