Figure 5.

Change of cytoplasmic pH in treated MCs using LysoSensor Green DND-189 staining. (a) MCs were seeded on coverslips in a 6-well plate and then were untreated (A, E, and I) or treated with VC (B, F, and J), MAP (C, G, and K), or AAE (D, H, and L) for 48 h. After treatment, the MCs were incubated with fresh medium containing 1 μM LysoSensor Green DND-189 for 30 min in the dark. The green fluorescence staining indicates the acidic cytoplasmic compartment. For pH neutralization, the MCs were simultaneously untreated (A–D) or treated with 10 nM Con A (E–H) or 10 mM NH₄Cl (I–L). Nuclei were stained with DAPI (blue). Scale bar: 10 μm. (b) Fluorescence intensity (arbitrary units (A.U.)) for LysoSensor Green DND-189 staining was measured using ImageJ. Histogram shows the results determined on 50 cells which are presented as means ± SD for three independent experiments. ^∗∗ P < 0.01 versus only VC or its derivatives.