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. 2020 Jun 30;9(7):1587. doi: 10.3390/cells9071587

Figure 4.

Figure 4

Effect of cAMP pathway regulation on apoptosis. Cells cultured in OM and treated with adenylate cyclase activator forskolin (FSK, 10 nM) and phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX, 500 nM) or with adenylate cyclase inhibitor SQ 22,536 (100 nM) for 72 h. Cells cultured in CM and OM with 10% DMSO were used as a reference for low and high apoptosis, respectively. Prior to flow-cytometry analysis, cells were labeled with Annexin V-FITC and 7-AAD. (a) Representative dot plots from flow cytometry analysis. (b) Mean population counts from three experiments performed on cells isolated from different donors; error bars represent standard deviation. Viable cells: Annexin V-/7-AAD–, early apoptotic cells: Annexin V+/7-AAD–, apoptotic/nonviable cells: Annexin V+/7-AAD+, necrotic/nonviable cells Annexin V-/7-AAD+. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, not significant; #, statistical significance of population count between marked experiment condition and CM sample (reference sample); number of # symbols denotes p-value as described above.