![Myo15 knock-down affects AZ scaffold, SVs, and EE. (A) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for RIM-BP (green) and HRP (magenta). (B–G) Quantifications of A. (B) RIM-BP spot area (control 100.0 ± 3.88% n = 12; Myo15-RNAi 85.07 ± 1.92%, n = 12). (C) Relative frequency histogram of RIM-BP spot areas (total number of spots control 3610 and Myo15-RNAi 4428), bin width 0.02. (D) Density of RIM-BP spots, as number of spots per NMJ area (control 100.0 ± 1.66% n = 11; Myo15-RNAi 109.8 ± 2.75%, n = 12). (E) RIM-BP mean pixel intensity of single spots (control 100.0 ± 9.63% n = 12; Myo15-RNAi 105.3 ± 3.39%, n = 12). (F) Total number of RIM-BP spots per NMJ (control 100.0 ± 2.82% n = 10; Myo15-RNAi 123.4 ± 5.68%, n = 12). (G) Total RIM-BP intensity as integral of the RIM-BP mean pixel intensity over the entire NMJ (control 100.0 ± 8.52% n = 12; Myo15-RNAi 125,5 ± 4.23%, n = 12). (H) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for VGlut (green) and HRP (magenta). (I) Quantifications of H. VGlut mean pixel intensity of NMJ (control 100.0 ± 3.85% n = 12; Myo15-RNAi 80.87 ± 2.22%, n = 12). (J) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for Fas2 (green) and HRP (magenta). (K) Quantifications of J. Fas2 mean pixel intensity of NMJ (control 100.0 ± 8.23% n = 12; Myo15-RNAi 113.6 ± 5.40%, n = 12). (L) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for Rbsn-5 (green) and HRP (magenta), only showing the NMJ contribution. (M–P) Quantifications of L. (M) Rbsn-5 (early endosome [EE]) spot area (control 100.0 ± 4.51% n = 11; Myo15-RNAi 84.47 ± 1.87%, n = 10). (N) Density of Rbsn-5 spots, as number of spots per NMJ area (control 100.0 ± 6.97% n = 11; Myo15-RNAi 139.8 ± 11.81%, n = 10). (O) Rbsn-5 (EE) mean pixel intensity of single spots (control 100.0 ± 10.3% n = 11; Myo15-RNAi 96.68 ± 13.63%, n = 10). (P) Total Rbsn-5 (EE) intensity as integral of the Rbsn-5 mean pixel intensity over the entire NMJ (control 100.0 ± 10.84% n = 10; Myo15-RNAi 130.0 ± 18.67%, n = 9). 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. VGlut, vesicular glutamate transporter 1; Fas2, fasciclin 2.](https://cdn.rupress.org/rup/content_public/journal/jcb/224/9/10.1083_jcb.202305059/1/jcb_202305059_fig2.png?Expires=1767639881&Signature=SPn~qqsL01qMQeyxm9BkgTnhmNstfr~jd6Y69SPZiLRSatuOArUe7Z2hdv7oy1z8i1IV4SfuGodHcBZ6GWZOGokDHq4kKo0BzrLewdjBt37E~6k1vbNpditcTWaTIIh8AU5tQZ1rp62auFKgeaTD8kmyNGIA~7YFp88uXCzxHQhpHaAU0t26bENkpx5ByWELZi8MEqqSzwH82ErDzMthQ1~gX1PxwSwsFBZReHRSpS7KMMceEgeFzQ7RUNIkZUkyB3XLXPmVWZUj9NurkbRgzAkIpdqLUPyvD~XOXt0aOiKDALsFq-IQpR-K~hNjKo5AcMbWHGLdB3gUSM7HukKsdw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Myo15 knock-down affects AZ scaffold, SVs, and EE. (A) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for RIM-BP (green) and HRP (magenta). (B–G) Quantifications of A. (B) RIM-BP spot area (control 100.0 ± 3.88% n = 12; Myo15-RNAi 85.07 ± 1.92%, n = 12). (C) Relative frequency histogram of RIM-BP spot areas (total number of spots control 3610 and Myo15-RNAi 4428), bin width 0.02. (D) Density of RIM-BP spots, as number of spots per NMJ area (control 100.0 ± 1.66% n = 11; Myo15-RNAi 109.8 ± 2.75%, n = 12). (E) RIM-BP mean pixel intensity of single spots (control 100.0 ± 9.63% n = 12; Myo15-RNAi 105.3 ± 3.39%, n = 12). (F) Total number of RIM-BP spots per NMJ (control 100.0 ± 2.82% n = 10; Myo15-RNAi 123.4 ± 5.68%, n = 12). (G) Total RIM-BP intensity as integral of the RIM-BP mean pixel intensity over the entire NMJ (control 100.0 ± 8.52% n = 12; Myo15-RNAi 125,5 ± 4.23%, n = 12). (H) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for VGlut (green) and HRP (magenta). (I) Quantifications of H. VGlut mean pixel intensity of NMJ (control 100.0 ± 3.85% n = 12; Myo15-RNAi 80.87 ± 2.22%, n = 12). (J) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for Fas2 (green) and HRP (magenta). (K) Quantifications of J. Fas2 mean pixel intensity of NMJ (control 100.0 ± 8.23% n = 12; Myo15-RNAi 113.6 ± 5.40%, n = 12). (L) Confocal images of RNAi-mediated Myo15 knock-down and control NMJs stained for Rbsn-5 (green) and HRP (magenta), only showing the NMJ contribution. (M–P) Quantifications of L. (M) Rbsn-5 (early endosome [EE]) spot area (control 100.0 ± 4.51% n = 11; Myo15-RNAi 84.47 ± 1.87%, n = 10). (N) Density of Rbsn-5 spots, as number of spots per NMJ area (control 100.0 ± 6.97% n = 11; Myo15-RNAi 139.8 ± 11.81%, n = 10). (O) Rbsn-5 (EE) mean pixel intensity of single spots (control 100.0 ± 10.3% n = 11; Myo15-RNAi 96.68 ± 13.63%, n = 10). (P) Total Rbsn-5 (EE) intensity as integral of the Rbsn-5 mean pixel intensity over the entire NMJ (control 100.0 ± 10.84% n = 10; Myo15-RNAi 130.0 ± 18.67%, n = 9). 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. VGlut, vesicular glutamate transporter 1; Fas2, fasciclin 2.