Fig. 1. Lyso-Gb3 induces autophagy in ARPE-19 cells.
(A) ARPE-19 cells were exposed to the indicated concentrations of lyso-Gb3 for 24 h. Transmission electron microscopic image of an autophagosome. Scale bar, 1 μm. (B) ARPE-19 cells were pretreated with 3-MA (10 mM) for 1 h and then incubated with lyso-Gb3 (0.5 μM) for 24 h. Autophagy was confirmed by indirect immunofluorescent staining using LC3 antibodies. Nuclei were stained with DAPI. Images were captured at an original magnification of ×600. Scale bar, 50 μm. (C) ARPE-19 cells were exposed to lyso-Gb3 (0.5 μM) for 24 h, and protein expressions were assessed by immunoblotting with anti-LC3B, anti-beclin-1, and anti-tubulin antibodies. Results are representative of three independent experiments. Bar graphs show densitometric quantifications of Western blot bands. (D) Cellular morphologies were observed under a light microscope using the same conditions. (E) Cell viabilities were determined using an MTT assay under the same conditions. Results are representative of three independent experiments. 3-MA, 3-Methyladenine. **p < 0.01 vs. vehicle controls, one-way ANOVA followed by Bonferroni’s post-hoc test.
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