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Fig. 6 Inhibiting exocytosis improves cartilage phenotypes in neu1 and gnptab deficient larvae (A) Schematic illustrates experimental workflow including histological stains, BMV109 labeling and analyses of heat shock induced Lamp1-mCherry transgene. (B) Live confocal analyses of Lamp1-mCherry (red) location in fli1a:EGFP positive neu1 deficient cartilages treated with either DMSO or 35 nM vacuolin. Boxed areas shown in red only in panels to the right. Percent values represent number of animals exhibiting the pictured phenotype. n = 10 larvae per condition. (C) Graph shows the percent of cells with cell surface localized Lamp1 in each condition. See Figure S4 for additional quantitation. Error = SEM, significance was assessed using an ANOVA, where ∗∗p < 0.01. (D) Alcian blue stained larvae show vacuolin treatment improves cartilage morphology in both neu1 and gnptab deficient animals. Percent values represent number of animals exhibiting the pictured phenotype. Scale bar: 20 μm. (E and F) Cartilage measurements in neu1 (E) and gnptab (F) morphants treated with either DMSO or vacuolin. n = 10–20 larvae per condition from 3 independent experiments. Error = SEM, significance was assessed using the Dunnet’s test (red stars), where ∗∗∗p < 0.001 and ∗∗∗∗p < 0.0001. (G) Alcian blue stained control and neu1 morphant larvae co-injected with lamp1a or lamp1b morpholinos show Lamp1 modulation improves phenotypes (see also Figure S4). Percent values represent number of embryos exhibiting the pictured phenotype. n = 10 larvae per condition. Scale bar: 20 μm. (H) Live confocal images of Lamp1-mCherry (red) in fli1a:EGFP (green) positive chondrocytes suggest Lamp1 modulation reduces cell surface lysosomal localization in neu1 morphants. (I) Quantitation of chondrocytes exhibiting cell surface localization. n = 15 larvae per condition. Error = SEM, significance was assessed using an ANOVA or the Dunnet’s test (red stars), where ∗∗∗∗p < 0.0001.