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. 2019 Jun 18;17:101–118. doi: 10.1016/j.isci.2019.06.020

Figure 7.

Figure 7

ANKHD1 Regulates the Size and Number of EEA1-Positive Early Endosomes by Vesiculation Ability

(A) Amounts of endogenous ANKHD1, endogenous EEA1, and expressed ANKHD1 in HeLa cells transfected with control siRNA + EGFP, ANKHD1 siRNA + EGFP, ANKHD1 siRNA + ANKHD1-EGFP, or ANKHD1 siRNA + ANKHD1-EGFP with 4E+ΔAH (4E+ΔAH-EGFP) for rescue experiments by western blot.

(B) Quantification of EEA1 in (A). Data represent the mean of four independent experiments.

(C) Confocal microscopic images of endogenous EEA1 for the rescue experiments as in (A). Scale bars, 10 μm.

(D–G) The total area (D) and the number (E), the size distribution (F), and the average area (G) of EEA1 puncta per cell in (C) were measured using ImageJ software. Data represent the mean of 27, 27, 25, and 25 cells from three independent experiments for control siRNA + EGFP, ANKHD1 siRNA + EGFP, ANKHD1 siRNA + ANKHD1-EGFP, and ANKHD1 siRNA + 4E+ΔAH-EGFP, respectively.

All error bars represent SE. *p < 0.05, **p < 0.01, and ***p < 0.001. Statistical significance was determined with the Student's t test. ns, not significant.