Figure 7.

A_2AR activation decreases cell surface deposition of LAMP2 in mouse IPMФs. (A) Immunofluorescence staining of IPMФ cells was made using LAMP2 specific, Alexa-488 conjugated antibody (green). The nuclei of macrophages were stained with DAPI (blue). LAMP2-specific fluorescence intensity was measured after LPS activation and treatment with the A_2AR agonist CGS21680 by Opera Phenix High Content Confocal System (Perkin Elmer, Waltham, MA, USA). Fifty fields and 500–1350 cells were acquired per well, and laser-based autofocus was performed at each imaging position. Images of DAPI and Alexa-488 channels were collected at 2 μm of the Z image plane using a 63× water immersion objective (NA: 1.15). Cellular features, such as the number of spots and relative spot intensities in the (B) membrane and (C) cytoplasmic regions, were extracted. Data obtained from the individual analysis of 500–1350 different cells are presented as mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001 control (vehicle-treated) vs. LPS activated cells and # p < 0.05; ### p < 0.001 LPS vs. LPS + A_2AR agonist-treated cells.