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Fig. 6 Analysis of autophagy pathway in cln8?/? mutants. (A) The graphs show the qRT-PCR analysis of autophagy factors, performed by normalizing to ?-actin, and comparing cln8?/? larvae with controls at 5 dpf. Three independent samples (each obtained from about 30?40 larvae) from controls and cln8?/? mutant larvae at 120 hpf were analyzed. Statistical analysis (* p ? 0.05, **p ? 0.01, *** p ? 0.001) was performed using Student's t-test. The values are expressed as mean ± standard deviation (SD). (B) Three independent larval homogenates from controls (n = 50) and cln8?/? larvae (n = 50) were tested by Western blotting for the expression of LC3, Rab7, Ctsd, and ATG5 protein. The levels of the proteins were normalized to ß-actin or GAPDH. Statistical analysis (*p ? 0.05, **p ? 0.01) was performed using Student's t-test. The values are expressed as mean ± standard error of the mean (SEM).