Fig. 2: LIN-2 and FRM-3 colocalize at neuromuscular junctions.
a Confocal detection of RFP-labeled GABAAR expressed from the unc-49::rfp knock-in allele kr296 and of GFP-LIN-2A expressed under the control of the muscle-specific promoter Pmyo-3. Fluorescence profiles indicate GFP-LIN-2A and RFP-UNC-49 fluorescence intensities along the nerve cord from the pictures above. b Confocal detection of RFP-LIN-2A and FRM-3A-GFP specifically expressed in muscle. The fluorescence profiles indicate RFP-LIN-2A and FRM-3A-GFP fluorescence intensities along the nerve cord from the pictures above. c In muscle cells, RFP-LIN-2A is present at GABAergic synapses labeled with NLG-1-GFP. d Pearson’s correlation coefficient between different markers. Box plots show lower and upper quartiles (box), median (center line) and standard deviation (whiskers). Kruskal–Wallis test (p < 0.001) followed by Dunn’s post test. ***p < 0.001; **p < 0.01; *p < 0.05. e–g Detection of surface-expressed GABAAR in the unc-49(kr378::pHluorin) knock-in strain shows small and diffuse clusters in the nlg-1(0) null mutant (f) as compared with the wild type (e); SNB-1-BFP labels GABAergic boutons (g) pHluorin-UNC-49 fluorescence levels were quantified and normalized to the wild type. Box plot as in Fig. 1e. Two-tailed Student’s t test ***p < 0.001. h–k Confocal detection of FRM-3B-GFP and GFP-LIN-2A expressed in muscle in wild-type animals or in nlg-1(ok259) mutants; SNB-1-BFP labels GABAergic boutons. j, k Quantification of FRM-3-GFP and GFP-LIN-2A fluorescence levels. Box plots are as in Fig. 1e. Two-tailed Student’s t test **p < 0.01. n.s: not significant. l Confocal detection of NLG-1-GFP expressed in the muscle of wild-type, frm-3(gk585) and lin-2(n397) animals. The fluorescence level of NLG-1-GFP was quantified and normalized to the wild type. Box plot as in Fig. 1e. One-way ANOVA followed by Tukey’s multiple comparison tests of each group compared with the wild type. *p < 0.05, n.s: not significant. Scale bars = 10 μm.
