Neuronal phenotype due to microtubule drug treatment or loss of klp-7, related to Fig. 1. (A) Confocal images of GFP-labeled (Pmec-7-GFP, muIs32) ALM neuron in untreated and 1 µM Taxol–treated worms. (B) The percentage of ALM neurons having ectopic growth due to treatment with different concentrations of Taxol. N = 3 independent replicates; n (number of neurons) = 65–142. (C) Representative images of PLM neurons in the untreated, 1 µM Taxol–treated, and 1 mM colchicine-treated worms showing the change in the length of the posterior process (double-sided arrow). Red arrowheads show ectopic branches. (D) Length of the posterior process of the PLM in worms grown in various concentrations of Taxol and colchicine. N = 3–5 independent replicates; n (number of neurons) = 45–95. (E) Confocal images of WT and klp-7(0) worms expressing Pmec-7-TagRFP::ELKS-1 (jsIs1075) + Pmec-7-GFP (muIs32). ELKS-1 puncta (magenta arrowheads) were noticed in the axonal and ectopic processes of the ALM. The white arrowheads indicate the enrichment of TagRFP::ELKS-1 at the synaptic region of PLM. (F) Confocal images of worms expressing Pmec-7-GFP (muIs32). Worms were immunostained using anti-GFP (shown in green) and anti–KLP-7 (shown in magenta) antibodies. The processed image to visualize the PLM-specific localization of KLP-7 is also presented separately, which is labeled as “α-KLP-7 (Only PLM).” (G) Confocal images and schematic of PVD neuron expressing Pdes-2::mCherry::RAB-3 (kyIs445) and PF49H124::GFP (wdIs52) reporters. Ectopic outgrowths (arrow) from the cell body and RAB-3 punctae (magenta arrowheads) in the dendrites were noticed in the klp-7(0) mutant. (H) The percentage of PVD neurons showing ectopic accumulation of mCherry::RAB-3 in dendrites. N = 4–5 independent replicates; n (number of neurons) = 25–30. Error bars represent SEM. Statistical comparison was done using the χ2 (Fisher’s exact) test for B and G. ANOVA with Tukey’s multiple comparison test was used for D. *, P < 0.05; ***, P < 0.001.
