Figure 3 | Scientific Reports

Figure 3

From: Cilostazol restores autophagy flux in bafilomycin A1-treated, cultured cortical astrocytes through lysosomal reacidification: roles of PKA, zinc and metallothionein 3

Figure 3The alternative text for this image may have been generated using AI.

Lysosomal free zinc levels are increased by treatment with cilostazol or cAMP. (a) Changes in intracellular and lysosomal free zinc levels were visualized by loading astrocytes with both FluoZin3-AM and LysoTracker for 30 minutes, followed by a sham wash (CTL) or treatment for 60 minutes with 10 μM cilostazol (n = 5) or 300 μM cAMP (n = 4). LysoTracker-positive particles exhibited red fluorescence in resting astrocytes (CTL). Cilostazol and cAMP further increased free zinc levels, most of which colocalized with LysoTracker fluorescence (Scale bar, 20 μm). (b) Changes in intracellular and lysosomal free zinc levels were visualized by loading astrocytes with a zinc-specific fluorescent dye Zinpyr-1 for 30 minutes, followed by a sham wash (CTL) or treatment for 60 minutes with 10 μM cilostazol, or 300 μM cAMP (n = 3) (Scale bar, 20 μm). (c) DND-189 fluorescence in astrocytes, before and after exposure to BafA1 alone (BA), BafA1 plus cilostazol (BA + Cilo), or BafA1 plus cAMP (BA + cAMP). Further addition of TPEN completely blocked the lysosomal pH-reversing effects of cilostazol (BA + Cilo+TPEN) and cAMP (BA + cAMP+TPEN) (Scale bar, 20 μm).

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