Fig. 4.

Effects of H₂O₂, CCCP, antimycin, and oligomycin on ATP depletion (ATP-dep). Pancreatic acinar cells were loaded with 4 μM Magnesium Green (MgGreen) for 30 min at room temperature to indirectly measure cytosolic ATP concentration. Representative traces show the relative MgGreen fluorescence (F/F₀; arbitrary units) in response to the “ATP depletion” cocktail with (gray trace) or without (black trace) preincubation with 10 μM BAPTA (A), 50 μM H₂O₂ (B), 500 μM H₂O₂ (C), 4 μM CCCP (D), 0.5 μM antimycin (E), and 10 μM oligomycin (F). The ATP depletion cocktail consisted of 100 μM carbachol (CCh), 10 μM oligomycin, and 2 mM iodoacetate and was used as a positive control to induce maximum ATP depletion. Cells that exhibited a rapid increase followed by a rapid decrease below the baseline fluorescence (see inset in A) were excluded from analysis because these likely represented cells undergoing cell lysis. Vertical bar represents 0.3 F/F₀ and horizontal bar represents 5 min. G: mean data were quantified and normalized by expressing the change in F/F₀ as a percentage of the ATP depletion cocktail response (*P < 0.05, as assessed using a one-sample t-test).