PrP-1 prevents the endocytosis and degradation of AJ components. a. E-cadherin (green) and β-catenin (magenta) localization in 6 hpf deep cells upon treatment with 10 µM Dynasore (DYNA); MO = lissamine-tagged PrP-1 morpholino (red); scale bar = 10 µm. b and d. Total levels of E-cadherin and β-catenin at 6 hpf upon treatment with Dynasore, MG-132 and Chloroquine (Chlq). c. Phenotypic analysis upon treatment with inhibitors. Top: progression of epiboly by 7.5 hpf is assessed by the downward extension of the embryonic margin (arrowheads) relative to the control (dashed horizontal line); brackets show the thickness of the blastoderm. Bottom: phenotypic quantification. Mean values are shown. e. AJ protein levels in 6 hpf embryos injected with increasing PrP-1 morpholino doses. WB = Western blot; IF = immunofluorescence. Red and black arrowheads in 1B, D and E indicate mature E-cadherin (120 kDa) and its more abundant precursor form (140 kDa), respectively. Densitometric analysis of Western blot bands is expressed in arbitrary units (AU) in B, D and E; average values of three independent experiments ± SEM are shown; statistical significance was assessed using unpaired, two-tailed t-tests; ns = not significant (p > 0.05), * = p ≤ 0.05, ** = p ≤ 0.01. In (e), E-cadherin and β-catenin levels were significantly reduced (p ≤ 0.05) at all three morpholino concentrations; p120ctn levels were not significantly changed at any morpholino dose (p > 0.05). See also Additional file 1: Figure S1.
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