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Fig. 7
Analysis of autophagy in epm2a−/− zebrafish mutants. (A) qRT–PCR analysis of autophagy factors (mtor, tfeb, beclin-1, atg5, atg12, lc3, and p62) normalized to β-actin in epm2a−/− larvae compared with controls at five dpf. Three independent RNA samples (each obtained from about 30–40 larvae) from controls and epm2a−/− mutant larvae at 120 hpf were analyzed. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, calculated by Student’s t-test. The values are expressed as mean ± standard deviation (SD). (B) Three independent larval homogenates from controls (n = 50) and epm2a−/− larvae (n = 50) were tested by Western blotting for the expression of ATG5, LC3–I and LC3–II proteins. The levels of the different proteins were normalized to the expression of β-actin. * p ≤ 0.05 was calculated by Student’s t-test.