Figure 8.

A_2AR activation modifies the localization of CtsD in LPS-activated macrophages.A, mouse IPMΦs were treated with CGS21680 (30 nM) in the absence or the presence of LPS (100 ng/ml) for 4 h. IS of mouse IPMΦs was conducted to determine the localization of endogenous CtsD protein (green) using CtsD-specific primary antibody and Alexa-488–conjugated secondary antibody. Nuclei of macrophages were stained with TO-PRO-3 iodide (red). Representative pictures are shown from 4 to 5 independent experiments. B, quantification of different parameters of the phagosomes from the immunostained IPMΦs was done by LAS X software (Leica Microsystem GmbH). The analysis was based on the individual densitometry of 50 different cells from 4 to 5 independent experiments. Data are presented as mean ± SD. Values from ANOVA: F = 52.44 and p < 0.001 for fluorescence intensity of and F = 9.022 and p = 0.0012 for average number of phagosomes. ∗p < 0.05 versus control (vehicle treated); ^###p < 0.001 versus LPS-activated cells. A_2AR, adenosine A_2A receptor; CtsD, cathepsin D; IPMΦ, mouse peritoneal macrophage cell; IS, immunostaining; LPS, lipopolysaccharide.