Fig. 2: Dystrophin and DAPC proteins determine TWK-28 content in the anterior domain of muscle cells. | Nature Communications

Fig. 2: Dystrophin and DAPC proteins determine TWK-28 content in the anterior domain of muscle cells.

From: Wnt-Ror-Dvl signalling and the dystrophin complex organize planar-polarized membrane compartments in C. elegans muscles

Fig. 2

A Confocal detection and quantification of mNeonGreen-TWK-28 in a long-isoform-specific dys-1 mutant (bln582) and a dys-1 molecular null allele (syb2174). n = 36, 37, 31 (for respective columns), N = 3 independent experiments. B Confocal detection and quantification of mNeonGreen-TWK-28 in dyb-1(cx36) and islo-1(bln549) loss-of-function mutants. n = 78, 64, 71 (for respective columns), N = 3 independent experiments. C C-terminal sequence of the TWK-28ΔPDZ variant engineered by gene editing of mNeonGreen-TWK-28 to eliminate a putative PDZ-binding motif (orange residues). D Confocal detection and quantification of mNeonGreen-TWK-28 in stn-1(ok292) and stn-2(ok2417) loss-of-function mutants, and of an mNeonGreen-TWK-28 knock-in line lacking the final four C-terminal amino acids (twk-28ΔPDZ). n = 38, 39, 32, 29 (for respective columns), N = 3 independent experiments. E Confocal detection and quantification of mNeonGreen-TWK-28 and mNeonGreen-twk-28ΔPDZ in a dys-1(syb2174) mutant background. n = 35, 33, 27, 33 (for respective columns), N = 3 independent experiments. F DYS-1 degradation protocol. L4 (P0) worms are transferred to auxin-containing media at day 0 (D0). F1 progeny are exposed to auxin until the L4 stage (ON Auxin) and transferred to NGM plates (OFF Auxin) for 24 h. Confocal detection and quantification of fluorescence at day 3 (D3) and 24 h later (D3 + 24 h). G DYS-1-AID-mNeonGreen is strongly degraded after lifelong exposure to auxin (ON Auxin, D3). Partial recovery 24 h after removal from auxin (OFF Auxin, D3 + 24 h). Scale bars, 20 µm. H, I Confocal detection of wrmScarlet-TWK-28 and DYS-1-AID-mNeonGreen at D3 and D3 + 24 h, in the absence of auxin (NO Auxin, age-matched controls), after auxin exposure for 3 days (ON Auxin), and after removal from auxin for 24 h (OFF Auxin). J Quantification of wrmScarlet-TWK-28 fluorescence shown in (H) and (I). n = 36, 50, 34, 43 (for respective columns), N = 3 independent experiments. Kruskal-Wallis test with Dunn’s multiple comparisons test; two-tailed Mann–Whitney test (for J). Each data point represents one muscle cell; bars represent data means; n.s., not significant, *p < 0.005. Raw data and statistical analyses are presented in the Source Data file. Image widths: 30 µm in A, B, D, E; 50 µm in (H, I).

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