Depletion of EB1, CLIP-170, or DCTN1 does not affect the distribution, uptake, or transport of various host cargoes. (A–D) NHDFs were transfected with control (Ctrl), EB1, or CLIP-170 siRNAs. (A) Cultures were stained for the early endosomal marker, EEA1 (green), Tyr-tubulin (blue), or nuclei using DRAQ5 (red). Representative widefield images are shown. (B) Quantification of mean distance of endosomes from the nucleus per cell in each sample (top) and fluorescence intensity of endosome staining (bottom; arbitrary units) demonstrated no significant differences between control and EB1 or CLIP-170 siRNA-treated samples (one-way ANOVA; distance: n = 1,380/group; F_(4137,2) = 0.281; P = 0.755; fluorescence intensity: n = 14/group; F_(39,2) = 1.187; P = 0.316). (C) Cultures were incubated for 30 min with fluorescently labeled transferrin (red). Processed samples were then stained for Tyr-tubulin (blue) and imaged. Representative widefield images are shown. (D) Transferrin uptake was determined by quantifying the fluorescence intensity of transferrin signal (arbitrary units) in samples described in C. No statistical difference was observed (one-way ANOVA; n = 20/group; F_(76,2) = 1.347; P = 0.265). (E) NHDFs were treated with control, EB1, CLIP-170, or DCTN1 siRNAs and then stained using MitoTracker or LysoTracker. Representative stills show mitochondria from Videos 2 and 3, or lysosomes from Videos 4 and 5. (F) NHDFs were treated with DMSO solvent control or 200-µM ciliobrevin D for 1 h and then stained using LysoTracker or MitoTracker. Representative still images show lysosomes from Video 6 or mitochondria from Video 7. Error bars represent standard error of the mean.