Fig.4.

Role of Kinesin-2α (KLP64D) tail in the anterograde axonal transport of Rab4-associated vesicles.

A, B) Synaptic FRAP profiles of Rab4-mRFP in the wild-type (WT), homozygous Klp64D^k5 (Klp64D^−/−), and Khc-73³⁶⁷³³ dsRNA expressing backgrounds. (A) and (B) depict a select set of time-lapse images in a pseudo color scheme (A), and the recovery of intensity values (average ± SD) with respect to the post-bleach intensities (B). Scale bar indicates 5 μm.

C) Schematic illustrates the composition of the heterotrimeric Kinesin-2 in Drosophila and mammals.

D–F) Localization of Rab4 in the ventral ganglion of third instar larvae in wild-type and homozygous Klp64D^k5 (Klp64D^−/−) backgrounds. Pseudocolored images of the representative ventral ganglia show relative levels of the endogenous Rab4 (D) and ectopically expressed Rab4-mRFP (E). Cell bodies show accumulated Rab4 in the cortex (white arrow) and decreased intensity in the neuropil (yellow circle) of the Klp64D mutant (E). The histograms depict relative enrichment (mean ± S.D.) of the endogenous Rab4 (magenta) and ectopically overexpressed Rab4-mRFP (indigo) in the wild-type and mutant backgrounds (F).

G) Kymographs of the Rab4-mRFP movement in lch5 axons. Rab4mRFP was expressed by chaGal4 in wild-type, the Klp64D^k5, and in two different transgenically rescued backgrounds (Klp64D^k5 Rescue - chaGal4>UAS-Klp64D-GFP/chaGal4>UAS-Rab4-mRFP; Klp64D^k5; and Klp64D^k5 ΔT-Rescue - chaGal4>UAS-Klp64DΔT-GFP/chaGal4>UAS-Rab4-mRFP; Klp64D^k5).

H–J) Traffic density (H), speed (I) and displacement (J) of Rab4-mRFP in lch5 axons. The p-values (*p<0.05, **p<0.01, ***p<0.001) were estimated using ANOVA.

(Movies S3 and S4A)