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Fig. 4. GLR-2 cytosolic tail sequences are sufficient to localize a heterologous membrane protein. (Top) The sequences of PES-10, GLR-1 and GLR-2 included in the TMGFP transgenes. (A) Full-length GLR-1::GFP is localized along the ventral nerve cord in a punctate pattern. (B) TMGFP alone and (C) TMGFP::GLR-1(tail) show no localization. (D) TMGFP::GLR-2(tail) has a similar clustered localization pattern (arrows) to full-length GLR-1::GFP. (A-D) Bars, 5 µm. (E,H) GLR-1::CFP. (F,I) GLR-2::YFP. (G) GLR-1::CFP and GLR-2::YFP colocalize at clusters (arrowheads) in neuron cell bodies (PVC is shown) and along proximal dendrites. (J) GLR-1::CFP and GLR-2::YFP colocalization also occurs at clusters (arrowheads) in the nerve ring (bracket) and along proximal dendrites of neurons in the head. (E-J) Bars, 10 µm. (K) GLR-1::YFP. (N,Q) TMYFP::GLR-2(tail). (L,O,R) SNB-1::CFP from PVD is localized to a small number of presynaptic boutons along the ventral cord neurites. (M) GLR-1::YFP colocalizes near SNB-1-labeled boutons (arrowheads). (P,S) Examples of two animals where TMYFP::GLR-2(tail) colocalizes with SNB-1-labeled boutons (arrowheads). (M,P,S) Merges from K and L, N and O, and Q and R, respectively. (K-S) Bars, 5 µm. All the transgenic animals shown here are of wild-type genetic backgrounds. The images were captured using FITC, CFP and YFP filters, and were taken from the ventral nerve cord region of the animal. (T) The mean number of clusters per 10 µm of ventral cord length, and (U) the mean cluster size is plotted for the given genotype. Error bars are s.e.m. for both graphs. **P<0.01 compared with GLR-1::GFP by ANOVA/Dunnett's multiple comparisons test. n=10-20 animals for each genotype.

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