Figure 8.
ANKHD1 Regulates the Size and Number of Rab5-Positive Organelles through Vesiculation Ability
(A) Amounts of endogenous ANKHD1 and expressed ANKHD1 in HeLa cells transfected with control siRNA + Venus×3, ANKHD1 siRNA + Venus×3, ANKHD1 siRNA + ANKHD1- Venus×3, or ANKHD1 siRNA + ANKHD1- Venus×3 with 4E+ΔAH (4E+ΔAH- Venus×3) for rescue experiments by western blot.
(B) Confocal microscopic images of expressed mCherry-Rab5Q79L in HeLa cells transfected with control siRNA + Venus×3 + mCherry-Rab5Q79L, ANKHD1 siRNA + Venus×3 + mCherry-Rab5Q79L, ANKHD1 siRNA + ANKHD1- Venus×3 + mCherry-Rab5Q79L, or ANKHD1 siRNA + 4E+ΔAH- Venus×3 + mCherry-Rab5Q79L for rescue experiments. Scale bars, 10 μm.
(C and D) Quantification of the number (C) and average area (D) of mCherry-Rab5Q79L puncta per cell in (B). Data represent the mean of 23, 19, 16, and 19 cells from three independent experiments for control siRNA + Venus×3, ANKHD1 siRNA + Venus×3, ANKHD1 siRNA + ANKHD1- Venus×3, and ANKHD1 siRNA + 4E+ΔAH- Venus×3, respectively.
All error bars represent SE. *p < 0.05, **p < 0.01, and ***p < 0.001. Statistical significance was determined using the Student's t test. ns, not significant.