FIGURE 3.

Co-localization of Pgp with EEA1-stained early endosomes increased after glucose variation-induced stress. Immunofluorescence microscopy images of KBV1 (+Pgp) (A) and KB31 (−Pgp) (B) cells incubated with low (0 mm), normal (25 mm), or high (50 mm) glucose for 2 h at 37 °C before staining with DAPI (blue) (i), EEA1 (red) (ii), and Pgp (green) (iii). Shown is overlay image of all three stains (iv) and a magnified image of the overlap (v). The overlap between EEA1 and Pgp is indicated by Mander's overlap co-efficient, R. DAPI, EEA1, and Pgp were quantified as fluorescence intensity/cell (vi). Pgp was also quantified as total Pgp fluorescence intensity co-localization with EEA1 using ImageJ software. *, p < 0.05; **, p < 0.01, and ***, p < 0.001 are relative to the respective 25 mm treated control glucose (e.g. 0 mm glucose KBV1 versus 25 mm glucose KBV1). Arrows indicate the co-localization of EEA1 with Pgp. Scale bar = 10 μm. ND stands for not detected.