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. 2018 Apr 19;9:242. doi: 10.3389/fphar.2018.00242

FIGURE 4.

FIGURE 4

Blocking P2Y12 reduces RPE autofluorescence in vitro. (A) Treatment of ARPE-19 cells with POSs and 10 μM chloroquine (CHQ) using the pulse chase method led to a substantial rise in autofluorescence at 488 nm ex/>525 nm em. FACS analysis demonstrates the increased autofluorescence with the CHQ/POS treatment for >6 days. Treatment of cells with 10 μM P2Y12 receptor antagonist AR-C66096 shifted the autofluorescence curve to the left. (B) Quantification of the mean autofluorescence from five independent FACS trials showing a significant reduction in POS/CHQ autofluorescence excited at 488 nm in cells treated with 10 μM AR-C66096 (#p = 0.02, ##p < 0.001, ∗p = 0.014, n = 5). (C) ARPE-19 cells displaying overlap of Magic Red and LysoTracker Green, suggesting cathepsin B activity in lysosomes under control conditions (top). Magic Red staining was eliminated by elevation of lysosomal pH with 10 μM tamoxifen (bottom). (D) Tamoxifen (10 μM) raises lysosomal pH in ARPE-19 cells (∗p = 0.012, n = 13).