Cortical dynein depends on recruitment from the cytoplasm by LIN-5 but not on EBP-2–mediated plus end tracking. (A) SDCM images showing eGFP::DHC-1 localization in control and lin-5(RNAi) metaphase embryos. Graphs show relative cortical over cytoplasmic eGFP::DHC-1 intensities as measured along the cortex (far right, green line) and normalized to a cytoplasmic line (red). Dotted lines at y = 100% indicate a 1:1 cortex/cytoplasm ratio. Mean (green) and individual (gray) traces are shown. n = 12 embryos and 24 cortexes for control, and n = 5 embryos and 10 cortexes for lin-5(RNAi). (B) SDCM images of anaphase embryos of indicated genotypes. Graphs show relative cortical over cytoplasmic eGFP::DHC-1 intensities, quantified and shown as described in A. n = 6 embryos per condition. (C) SDCM images show mCherry::DHC-1 and eGFP::LIN-5 localization. Mean (thick lines) and individual (thin lines) intensity plots are shown in left graphs. Right graphs show cross-correlation between cortical LIN-5 and DHC-1 plots expressed as means ± SEM. n = 10 embryos and 20 cortexes. (D) SDCM images showing dynein or LIN-5 localization in perm-1(RNAi) or perm-1(RNAi); lin-5(RNAi) embryos treated with 1 µM nocodazole (noc.). Graphs indicate fluorescence intensity profiles as measured in dashed boxes shown in superimposed panels. The red arrow points to mitotic spindle remnant. Plots are mean (n = 10) intensities relative to cytoplasmic values ± SD. Bars, 5 µm.