Migfilin primarily localizes at late endosomes/lysosomes. KP4 cells were stably transfected with mCherry-Migfilin and DFCP1–GFP or GFP-LC3, GFP-Rab7 under control or EBSS starvation condition (Starv.), and then imaged by fluorescence microscopy. (A–D) Representative images were shown in the left panel and quantification analysis was shown in the right panel. Scale bar: 10 µm (Magnify: 2 µm). The colocalization ratio of mCherry-Migfilin with DFCP1-GFP (A), GFP-LC3 (B), GFP-Rab7 (C) or Lysotracker Green (Lyso-Green) (D) was determined. At least 18 cells in each group were analyzed. Data represent mean ± SE, *P < 0.05, n.s., no significance, unpaired two-tailed t test. n = 3 independent experiments.