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. 2024 May 10;13(5):bio060338. doi: 10.1242/bio.060338

Fig. 1.

Fig. 1.

In ADAP1 and ARAP1KD cells, CD63 incorporation in endosomes is inhibited. (A) MCF7 cells were transfected with control and ADAP1 siRNAs, and the cell lysates were subjected to western blotting. Note that ADAP1 was efficiently downregulated in ADAP1 siRNA-transfected cells compared with control siRNA-transfected cells. (B) A similar experiment for ARAP1. Note that ARAP1 was efficiently downregulated by its siRNA. (C) The siRNA-transfected cells were transfected with GFP-Rab5Q79L (green) and stained with anti-CD63 (red). The insets were enlarged and Rab5 positive endosomes are indicated with arrowheads. Note that CD63 was filled in endosomes in control cells, whereas CD63 signal was lost in ADAP1 and ARAP1KD cells. (D) The Rab5-positive endosomes were classified 100% filled, partially filled, and none with CD63. The percentage of Rab5 endosomes in each category per cell was shown. More than 15 cells were counted per experiment and the experiment was repeated four times. Error bar, standard deviation (s.d.). Control, n=91; ADAP1KD, n=73; ARAP1KD, n=67. Significance was calculated by two-way ANOVA with Sidak's multiple comparisons test. ns, not significant, ****P<0.0001. Scale bars: 10 µm.