FIG. 1.

Accumulation of autophagosomes in epithelial cells treated with artificial tears containing sodium hyaluronate and trehalose. (A, B) GFP-LC3 HeLa cells were treated with Hyabak or Thealose Duo for 4–24 h. Treatment with trehalose alone at a final concentration of 40 mM was used as a known inducer of autophagy. Nuclei were stained with DAPI. Scale bar = 10 μm (A) Representative images. (B) Quantification of the number of GFP-LC3 dots per cell. (C, D) GFP-LC3 HeLa cells were treated with Hyabak or Thealose Duo during 24 h. Treatment with EBSS (nutrient starvation) was used as a known inducer of autophagy. (C) Representative images of 3 independent experiments. Scale bar = 10 μm. (D) Quantification of the number of dots per cell. *p < 0.05; ***p < 0.001; ****p < 0.0001 (one-way Anova test) compared with the untreated control cells. EBSS, Earle's balanced salt solution; GFP, green fluorescent protein.