Fig. 4 | Cellular and Molecular Life Sciences

Fig. 4

From: Bafilomycin A1 activates respiration of neuronal cells via uncoupling associated with flickering depolarization of mitochondria

Fig. 4

Baf reduces ΔpHm and ΔΨm in dPC12 cells. a Inhibition of the F0F1 ATPase with oligomycin (10 μM) decreases Baf-specific cell deoxygenation by 10–20%. b Oligomycin treatment significantly elevates the fluorescence of the mtAlpHi probe in control cells, indicating a sustained matrix alkalinization. In Baf(+) cells the increase in ΔpH is lower and not sustainable. c TMRM profiles show a gradual decrease in probe fluorescence in Baf(+) cells upon addition of oligomycin. d Uncoupling with 0.5 μM FCCP causes a drop in TMRM and mtAlpHi fluorescence in Baf(+) cells, demonstrating a decrease in the ΔΨm and ΔpH, respectively (F0/F represents the ratio of fluorescence before and 3 min after FCCP addition). The decrease in mtAlpHi signal does not depend on Baf treatment; in contrast, the relative drop in TMRM signal in Baf(+) cells is significantly smaller than in the control. In all experiments except that shown in a, cells were preincubated with 0.25 μM Baf for 45 min prior to the addition of oligomycin or FCCP. DMSO was used as a negative control. Bars 20 μm; asterisks significant differences

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