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. Author manuscript; available in PMC: 2022 Apr 7.
Published in final edited form as: Ocul Surf. 2021 Jun 11;21:193–205. doi: 10.1016/j.jtos.2021.06.006

Fig. 3.

Fig. 3.

EphA2 regulates autophagy: (A) Venn diagram showing the overlap of potential EphA2 interacting proteins and autophagy genes (GO consortium, GO:0006914). (B) Phase-contrast microscopy of hTCEpi cells 48hr after siControl or siEphA2 transfection. (C) Number of cells with vacuoles were counted in (B). Three fields (299 μm × 237 μm) per well in each experiment were counted. Each experimental replicates included 3 technical replicates. Scattered dots: values for replica. (D) Confocal imaging of LAMP1 (red) and LC3 (green) immunofluorescent staining in hTCEpi cells after siControl or siEphA2 transfection. White arrows: LC3 puncta on the membranes of LAMP + vesicles. (E–F) Western blotting showing EphA2, p62, LC3I and LC3II levels in hTCEpi cells after siControl or siEphA2 transfection (E) as well as in mouse corneal epithelium isolated from EphA2 KO and littermate control mice (F; 6 weeks old). Graphs indicate the densitometry analysis of blots for LC3II and p62 levels. * = p < 0.05. GAPDH was used as a loading control. (G–H) Fluorescence images showing GFP-LC3 puncta in EphA2 KO mice and littermate controls (WT) corneal epithelium. DAPI was used to stain nuclei. White dotted lines demarcate the basement membrane region. The GFP-LC3 signal in a region of wing cells in EphA2 KO cornea was enhanced to show the puncta. Number of LC3 puncta was counted using ImageJ (H). Scattered dots: values for an individual mouse.