Figure 1
The A2AR agonist CGS21680 rescues the effects of U18666a on viability, morphology and cholesterol accumulation. (A) Cell viability was assessed by MTT reduction assay in OPs, at 3 DIV, treated for 48 h with 1.25 μM of U18666a (U), 100 nM of CGS21680 (CGS) and 100 nM of ZM241385 (ZM). Data are presented as mean percentages of control cultures ± SEM, n = 4–6 independent experiments run in triplicate (*p < 0.001 vs CTR **p < 0.01 vs U ***p < 0.05 vs U + CGS). (B) Phase contrast morphology of cultured OPs treated for 48 h with U18666a and CGS21680 were shown. Scale bar = 50 µm (C–E). Cholesterol was labeled with Filipin III (blue). (C) The OPs were treated for 48 h with U18666a alone or with CGS21680. CGS21680 was added together with U18666a or in the last 24 h of treatment. Mean fluorescence intensities (MFI value) of Filipin III are shown in D. Data are mean ± SEM, n = 150 cells for condition (*p < 0.001 vs CTR **p < 0.001 vs U). (E) Filipin III was evaluated in cultures treated with A2AR antagonist ZM241385 (100 nM), added 30 min earlier than the other drugs. (F) A2AR expression in OP, at 1 and 2 DIV, treated or not for 24 h with U18666a at 1DIV. (G) Mean fluorescence intensities (MFI value) were shown. Data are mean ± SEM of n. 150 cells for condition (*p < 0.005 vs CTR. Scale bar = 50 µm).
