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Fig. 3 Inactivation of Vcp leads to an accumulation of ubiquitinated proteins and Sqstm1. (a) Western blot analysis using an anti-K48-linkage specific ubiquitin antibody after MO1-control and MO1-vcp injection and DMSO or MG132 treatment (n = 4). (b) Vcp morphants show a significant increase of Sqstm1 by western blot analysis (n = 3); quantification of gray values. Actb1/β-actin was used as loading control. Data represent means ± SD, unpaired Student t test, *P value< 0.03 (c) Western blot analysis of Lc3 after MO1-control and MO1-vcp injection and DMSO or Ammonium chloride (NH4Cl) treatment (top) and quantification of gray values (bottom). NH4Cl treated control embryos reveal significant increase of Lc3-II levels, compared to DMSO treated embryos (n = 5). Actb1/β-actin was used as loading control. The individual samples are displayed (two-sided Wilcoxon rank-sum test).