Journal: The Journal of Cell Biology
Article Title: Myosin 15 participates in assembly and remodeling of the presynapse
doi: 10.1083/jcb.202305059
Figure Lengend Snippet: Analysis of AZ, SV, endocytic and cell adhesion proteins, and mitochondria upon motoneuronal Myo15 knock-down. (A) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for liprin-α (green) and HRP (magenta). (B) Quantifications of A. Liprin-α mean pixel intensity of NMJ (control 100.0 ± 8.04% n = 11; Myo15-RNAi 89.1 ± 8.38%, n = 12). (C) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for Rab3 (green) and HRP (magenta). (D) Quantifications of C. Rab3 mean pixel intensity of NMJ (control 100.0 ± 2.83% n = 12; Myo15-RNAi 82.0 ± 2.36%, n = 12). (E) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for DAP160 (green) and HRP (magenta). (F) Quantifications of E. DAP160 mean pixel intensity of NMJ (control 100.0 ± 11.0% n = 9; Myo15-RNAi 89.95 ± 6.04%, n = 10). (G) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for Nrg (green) and HRP (magenta). (H) Quantifications of G. Nrg mean pixel intensity of NMJ (control 100.0 ± 5.51% n = 11; Myo15-RNAi 105.0 ± 9.85%, n = 10). (I and J) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for ATP5 (mitochondria) (green) and HRP (magenta) with NMJ and muscle contribution, and (J) only NMJ ATP5 contribution. (K–N) Quantifications of J. (K) ATP5 spot (mitochondria) area (control 100.0 ± 5.13% n = 12; Myo15-RNAi 103.2 ± 5.22%, n = 12). (L) Density of ATP5 spots (mitochondria) as number of spots per NMJ area (control 100.0 ± 5.92% n = 12; Myo15-RNAi 97.35 ± 5.02%, n = 12). (M) ATP5 (mitochondria) mean pixel intensity of single spots (control 100.0 ± 11.67% n = 12; Myo15-RNAi 91.4 ± 10.06%, n = 12). (N) Total ATP5 (mitochondria) intensity as integral of the ATP5 mean pixel intensity over the entire NMJ (control 100.0 ± 11.04% n = 12; Myo15-RNAi 101.4 ± 11.71%, n = 10). (O) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for Rbsn-5 (EE) (green) and HRP (magenta) with NMJ and muscle contribution. For all images, scale bar overview 5 µm, zoom 2 µm. All data are provided as mean ± SEM. N represents NMJs with 1–2 NMJs/animal from five to six animals, *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Nrg, neuroglian.
Article Snippet: Neuroglian (Nrg, anti-mouse, 1:50) , Developmental Studies Hybridoma Bank , BP 104.
Techniques: Knockdown, Control, Staining
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