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Figure 7


Fig. 7. Involvement of actin cytoskeleton in vps35 mutant phenotypes. (A) Labelling of F-actin in WT and vps35 haemocytes, visualised using single confocal sections as in Fig. 4E. Arrow indicates plasma membrane surrounding the body of the cell. Arrowhead indicates leading edge of lamellopodia. (B) F-actin levels in WT, vps35, vps35/Df, vps35/Df;Hml-Gal4 and vps35/Df; Rac1/+ haemocytes and in vps35 haemocytes treated with 0.1 mM cytochalasin D (cytoD) (average fluorescence per cell). n=25-30 cells from four larvae; statistical comparisons are with vps35/Df. (C) Removal of a single copy of Rac1, or treatment with 0.1 mM cytochalasin D, partially rescues the loss of mBSA-Texas-Red uptake in haemocytes that lack Vps35. n=25-30 cells from four larvae. (D) The vps/Df supernumerary bouton phenotype is suppressed by removal of a single copy of Rac1, or by presynaptic or postsynaptic expression of dominant-negative Rac1 (UAS-Rac1.N17). Note that elav-GAL4 and BG57-GAL4 also drive Vps35 expression and partially rescue the vps35 mutant phenotype (see Fig. 6B). NMJs from muscles 6 and 7 in abdominal segment 3 were used for all analyses; n=3-8 larvae. All statistical comparisons are with WT except where indicated. All data are mean ± s.e.m.





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