,
, BUT INDEPENDENTLY OF EBP
Supplemental Figure S5C,D), we set out to confirm that that dynein uses EBP-2 EB to track microtubule plus ends, as previously reported, Because DYCI-1::mCherry displayed similar dynamics to EBP-2::GFP and colocalized with it, 2017. ,
To further test whether dynein hitches a ride with EBP-2, either directly or with the help of accessory proteins, we depleted dynactin and LIS-1 by dnc-1 p150/glued (RNAi) and lis-1(RNAi), respectively. These RNAi were partial, to preserve the early steps of mitosis. We observed a strong reduction in the LSP-directed track densities, suggest that in higher eukaryotes dynein tracks the microtubule plus ends via a hierarchical interaction involving binding to dynactin, which in turn binds to EB1 with the help of CLIP170, 1998. ,
We concluded that dynein accumulates at the microtubule plus ends via EBP-2 EB with the help of dynactin and LIS-1, which resembles the findings in mammal protein experiments, 2014. ,
We tracked spots on both channels, setting as colocalized those spots that are closer than 4 pixels at each time. We studied only those tracks longer than 6 time-points and which displayed directed motion, although we reduced the threshold track length to 3 points for cortical colocalizations. In all cases, we wondered whether the high density of DYCI-1::mCherry spots might cause artefactual colocalization. Therefore, for each colocalization experiment, we compared the results with the colocalization of a synthetic set of spots of identical count, We crossed the strain carrying randomly integrated DYCI-1::mCherry with ones carrying either TBA-2::YFP or EBP-2::GFP. We found that most of the DYCI-1::mCherry spots colocalized with microtubule plus ends and EBP-2::GFP in the doubly labeled strains (Supplemental Figure S5A-D and Movie S10-11), 2011. ,
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