Fig. 5: DG glycosylation is decreased by the absence of DNRX in Drosophila muscle. | Communications Biology

Fig. 5: DG glycosylation is decreased by the absence of DNRX in Drosophila muscle.

From: Neurexin facilitates glycosylation of Dystroglycan to sustain muscle architecture and function in Drosophila

Fig. 5

a Decrease in DG glycosylation in the absence of DNRX. DG was labeled with an anti-HA antibody, and DG levels were subsequently analyzed via lectin blotting and western blotting. b Quantification of DG glycosylation via lectin blotting from the data in (a). The data are presented as the means ± SEMs of three independent experiments. **p < 0.01, *p < 0.05; two-tailed unpaired Student’s t-test. c Western blotting analysis that DNRX and increased DG glycosylation were present following density gradient centrifugation. The samples were separated into 19 fractions with varying densities through centrifugation, and the fractions were labeled 1 to 19. Bip was used as an ER marker. Syx16 was used as a Golgi marker. H: Heavily glycosylated DG, L: Lightly glycosylated DG. d The grayscale values of the bands corresponding to DNRX, DG-H and DG-L in each lane. e Quantification of the ratio of grayscale values of the DG-H to DG-L bands in lanes 9 to 17. The data are presented as the means ± SEMs of three independent experiments. f Western blot analysis revealed that loss of DNRX decreased DG glycosylation in the microsomal fraction. g Quantification of the ratio of DG-H to DG-L in the microsomal fraction. The data are presented as the means ± SEMs of three independent experiments. **p < 0.01; two-tailed unpaired Student’s t-test. h Western blot analysis revealed a decrease in DG glycosylation on the cell membrane in the absence of DNRX. MF: membrane fraction, CF: cytoplasmic fraction. An anti-HA antibody was used to label DG, ATP-α was used as a membrane fraction marker, and GAPDH was used as a cytoplasmic fraction marker. i Quantification of the ratio of DG-H to DG-L on the plasma membrane. The data are presented as the means ± SEMs of four independent experiments. ***p < 0.001; two-tailed unpaired Student’s t-test. j Western blot analysis revealed that DG glycosylation was rescued by the overexpression of full-length DNRX in muscle. Tubulin was used as a cytoplasmic fraction marker. k Quantification of the ratio of DG-H to DG-L on the plasma membrane. The data are presented as the means ± SEMs of three independent experiments. *p < 0.05, two-tailed unpaired Student’s t-test.

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