(A) FRM-3′s ERM domain binds LIN-2A in yeast 2-hybrid assays. Growth of Y2HGold cells on selective media (–Trp/-Leu/-His/-Ade) is shown. Yeast cells were transformed with vectors expressing the indicated fusion proteins. Positive (+, pGBKT7-53 and pGADT7-T) and negative (−, pGBKT7-Lam and pGADT7-T) controls are indicated. ERM domains derived from FRM-1, FRM-2 and FRM-3 were tested for interaction with LIN-2A. (B) Muscle expressed GFP-UNC-49B (Green) and LIN-2::mCherry (Red) are co-localized in the nerve cord. A representative image is shown (scale bar 5 μm). (C, D) GFP-UNC-49B puncta fluorescence in the nerve cord was decreased in lin-2 mutants. This defect was rescued by transgenes expressing LIN-2A in body muscles (M) but not by those expressed in GABergic neurons (N). Representative images (C, scale bar 5 μm) and mean puncta intensity (D) are shown. (E–G) mIPSC amplitude was reduced in lin-2 mutants and this defect was rescued by a transgene expressing LIN-2 in body muscle (M resc). mIPSCs were recorded from adult body muscles. Representative traces (E), mean amplitude (F), and mean frequency (G) are shown. (H) Muscimol-activated currents in adult body muscles were unaffected in lin-2 mutants, indicating that the function of total surface UNC-49 receptors was unaltered. Mean peak currents are shown. lin-2 and frm-3 mutations did not have additive effects on UNC-49B puncta fluorescence (D) or mIPSC amplitudes (F) in double mutants. Values that differ significantly are indicated (***, p < 0.001; **, p < 0.01; ns, not significant). The number of animals analysed is indicated for each genotype. Error bars indicate SEM.