Ppk1 localizes to axons when dynein-mediated transport is disrupted.

Representative Images of ddaC neurons in live 3rd instar larvae. Dashed-outline boxes: Zoomed-in views of axons. (A) Representative images and quantification of Ppk1::sfGFP in control (21 neurons, 11 larvae) and Dlic-RNAi-expressing neurons (21 neurons, 11 larvae). Quantification, axons: Mann-Whitney test (pdmn (O/E dmn). Quantification, axons: Mann-Whitney test (pO/E dmn (23 neurons, 14 larvae). Scale bars, 50 μm and 5 μm (dashed-outline boxes). (B) Representative images of Ppk1::sfGFP and GALNT2::TagRFP in the axons of control and Dlic-RNAi-expressing neurons. Zoomed in images of an axon of a Dlic-RNAi-expressing neuron shows that the Ppk1::sfGFP puncta (green arrowheads) do not colocalize with ectopic Golgi marked by GALNT2::TagRFP (magenta arrowheads). Scale bars, 10 μm and 5 μm (dashed-outline boxes). (C) Representative images of GALNT2::TagRFP and Ppk1::sfGFP in the axons of control neurons and neurons expressing a dominant-negative form of the Golgin Lava lamp (Lva) (Lva-DN); Lva-DN disrupts the interaction between Golgi and dynein but does not affect dynein activity. In neurons expressing lva-DN, GALNT2::TagRFP mislocalizes to axons (black arrowheads) and axonal Ppk1::sfGFP levels increase. Quantification, Ppk1::sfGFP, axons: Student’s unpaired t-test (plva-DN (16 neurons, 7 larvae). Scale bar, 10 μm. Control genotypes: w1118; ppk-Gal4. Experimental genotypes: w1118; ppk-Gal4 UAS-Dlic-RNAi UAS-Dicer (A and B), w1118; ppk-Gal4 UAS-dmn (A), w1118; ppk-Gal4 UAS-lva-DN (C). UAS-GALNT2::TagRFP and ppk1::sfGFP included as indicated (A-C). In the graphs, each data point represents a neuron and data are plotted as mean ± SEM. ****p (PDF)