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. 2018 Sep 3;20(10):1008–1022. doi: 10.1016/j.neo.2018.08.005

Figure 2.

Figure 2

Knockdown of AXL expression decreases peripheral distribution of lysosomes in EAC cells. (A-D) LAMP1 IF analysis. (A) Representative confocal IF images (60×) of LAMP1 (green) in SK-GT-4-shControl versus SK-GT-4-shAXL cells, or (C) FLO-1-shControl versus FLO-1-shAXL cells. Arrows point to cells with peripheral lysosomes, while arrowheads indicate cells with perinuclear lysosomes. (B) Quantification of the percentage of SK-GT-4-shControl versus SK-GT-4-shAXL cells, or (D) FLO-1-shControl versus FLO-1-shAXL cells with either peripheral or perinuclear lysosomes. Data are represented as mean ± SE. (E-H) Live-TIRF microscopy analysis of lysosomes peripheral localization. (E) Representative TIRFM images (60X) of Lysotracker (+) acidic vesicles in SK-GT-4-shControl versus SK-GT-4-shAXL cells, or (G) FLO-1-shControl versus FLO-1-shAXL cells. (F) Quantification of total number of tracks per cell present close to the plasma membrane of SK-GT-4-shControl versus SK-GT-4-shAXL cells, or (H) FLO-1-shControl versus FLO-1-shAXL cells. Data are represented as median ± SD.