CME is required for Rab35 and Rab5/Rab11 endosomal dynamics. (A and B) Still frames of embryos expressing YFP:Rab11 (A) and YFP:Rab5 (B) injected with water control or chlorpromazine (chlor). (C) Quantification of the number of Rab5 and Rab11 compartments when injected with water control and chlorpromazine. n = 132 (Rab5, control), 108 (Rab5, chlor), 118 (Rab11, control), and 124 (Rab11, chlor) cells. (D) Quantification of the size of Rab5 and Rab11 when injected with water control and chlorpromazine. n = 284 (Rab5, control), 280 (Rab5, chlor), 258 (Rab11, control), and 350 (Rab11, chlor) compartments. (E) Distribution of Rab11 compartmental size in control and chlorpromazine injected embryos. (F and G) Images of the association between YFP:Rab5 and mCh:Rab35 in control (F) and chlorpromazine injected embryos (G). Arrowheads mark associated Rab5 and Rab35 compartments. Rab5 and Rab11 localization is largely unaffected by coexpression of mCh:Rab35 (Fig. S5, A–D). (H) Distribution of Rab35 and Rab5 normalized association times in control and chlorpromazine injected embryos. The association time was measured and normalized to each compartmental lifetime. n = 105 (control) and 101 (chlor) compartments. (I) Quantification of the colocalization among Rab35, Rab11, and Rab5 in control and chlorpromazine-injected embryos. n = 113 (Rab5, control), 160 (Rab5, chlor), 144 (Rab11, control), and 107 (Rab11, chlor) compartments. Scale bars in A, B, and F represent 5 μm. In bar plots, error bars indicate standard errors. Statistical significance was calculated using a Mann–Whitney U test (C and D) and Student’s t test (I). *, P < 0.05; ***, P < 0.0005. In A, B, F, and G, embryos are oriented with anterior up and posterior down.