In the absence of Lis1, dynein fails to colocalize with endosomes or peroxisomes at the hyphal tip. (A–D) Micrographs of the extreme tips of wild-type (A and C) and Lis1Δ (B and D) hyphae expressing dynein-3xGFP and mCherry-Rab5/RabA–labeled endosomes (A and B) or mCherry-PTS1–labeled peroxisomes (C and D). In wild-type hyphae, endosomes (A) and peroxisomes (C) localize along the length of microtubules, labeled by dynein-3xGFP, which decorates the microtubule plus ends and localizes to discrete puncta. Dynein-3xGFP colocalized with moving endosomes and peroxisomes (labeled with white asterisks in A and C), as determined by kymograph analysis. In Lis1Δ hyphae, dynein-3xGFP localizes to comets at the microtubule plus ends and to the cytoplasm (B and D, top) but does not colocalize with either endosomes (B, bottom) or peroxisomes (D, bottom) accumulated at the hyphal tip. Dashed lines indicate the outline of the hyphae. Bars, 1 µm.