Figure 6.

Neuronal expression of a constitutive active form of Trc (Trc^Myr) rescues the dendritic tiling defects of the sin1 and rictor mutants. (A) S2 cells transfected with wild-type Trc (Trc-Flag), a membrane-targeted Trc (Trc^Myr-Flag), or a membrane-targeted Trc with a mutation at the Thr449 site (Trc^Myr(T449A)-Flag) were stained with anti-Flag antibodies. Images were taken under a confocal microscope. Scale bar=10 μm. (B) Lysates of S2 cells expressing a Flag-tagged wild-type (WT) (Trc-Flag), a membrane-targeted Trc (Trc^Myr-Flag), a membrane-targeted Trc with a mutation at the Thr449 site (Trc^Myr(T449A)-Flag), or a membrane-targeted Trc with a kinase-dead mutation at the Lys122 site (Trc^Myr (K122A)-Flag) were analyzed by blotting using anti-Flag (top panel) and anti-Thr449P (bottom panel) antibodies. (C–H) In sin1 and rictor mutant larvae carrying a single copy of UAS-trc^Myr under the control of ppk-Gal4 driver, tiling defects in ddaC dendrites were largely rescued. Scale bar=50 μm. Genotypes: (C) yw; sin1^PBac/sin1^PBac; ppk-EGFP/ppk-EGFP, (D) yw; UAS-trc, sin1^PBac/sin1^PBac; ppk-Gal4, ppk-EGFP/ppk-EGFP, (E) yw; sin1^PBac, UAS-trc^Myr/sin1^PBac; ppk-Gal4, ppk-EGFP/ppk-EGFP, (F) yw, rictor^Δ2/yw, rictor^Δ2; +/+; ppk-EGFP/ppk-EGFP, (G) yw, rictor^Δ2/yw, rictor^Δ2; UAS-trc/+; ppk-Gal4, ppk-EGFP/ppk-EGFP, and (H) yw, rictor^Δ2/yw, rictor^Δ2; UAS-trc^Myr/+; ppk-Gal4, ppk-EGFP/ppk-EGFP. (I, J) Quantification of the dendritic crossing points (I) and the total branch length (J) in the rescue experiments. −, sin1 or rictor mutants carrying no transgene. Error bars indicate the mean±s.d. (n=15), ^*P<0.01 (Student's t-test).