Journal: The Journal of Biological Chemistry

Article Title: Cathepsin D interacts with adenosine A 2A receptors in mouse macrophages to modulate cell surface localization and inflammatory signaling

doi: 10.1016/j.jbc.2022.101888

Figure Lengend Snippet: A 2A R 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 2A R, adenosine A 2A receptor; CtsD, cathepsin D; IPMΦ, mouse peritoneal macrophage cell; IS, immunostaining; LPS, lipopolysaccharide.

Article Snippet: Densitometry analysis was completed by Leica LAS X Life Science software version.

Techniques: Activation Assay, Staining, Software, Fluorescence, Immunostaining