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Gene Synthesis> | Plasmid and Drosophila line construction To generate Axin targeted to mitochondria, the region that encodes the short C-terminal ActA mitochondrial targeting sequence [66] was synthesized by Genscript and cloned via Clone EZ Technology downstream of tDimer-Red12 (RFP) [90] in pUAST to create a carboxyl-terminal fusion. | Get A Quote |
Dendrite microtubules are polarized with minus-end-out orientation in Drosophila neurons. Nucleation sites concentrate at dendrite branch points, but how they localize is not known. Using Drosophila, we found that canonical Wnt signaling proteins regulate localization of the core nucleation protein γTubulin (γTub). Reduction of frizzleds (fz), arrow (low-density lipoprotein receptor-related protein [LRP] 5/6), dishevelled (dsh), casein kinase Iγ, G proteins, and Axin reduced γTub-green fluorescent protein (GFP) at branch points, and two functional readouts of dendritic nucleation confirmed a role for Wnt signaling proteins. Both dsh and Axin localized to branch points, with dsh upstream of Axin. Moreover, t... More