ATG10 is involved in p62-regulated autophagy.

(A) Scatter plots comparing the normalized, relative expression of autophagy-related proteins in ARPE-19 cells transfected with p62 specific or scrambled siRNA (10nM) for 24 h. mRNA expression was detected using autophagy qRT-PCR array from SA Biosciences each. The 'fold-change boundary' denoted by the parallel lines flanking the median line was set at 2 and segregates the genes up or down regulated. Dots represent genes plotted on the basis of Log ₁₀(2^-ΔΔCt). Blue circles denote the genes ATG10 and p62 with -2.44 and -6.77 fold changes respectively. (B) p62 mRNA level was examined by qRT-PCR to confirm the knockdown of p62 transcription and mRNA level of autophagic protein ATG10 was assessed in p62-knockdown cells. (C) RPE cells were treated by 400μM H₂O₂ and ATG10 mRNA level was measured by qRT-PCR. Mean+ S.E.M. from three independent experiments were plotted. (D) RPE cells were transfected with p62 or scrambled siRNA for 24h and then exposed with 400 H₂O₂ for 1, 3, 6, 12h respectively. Whole-cells lysates were analyzed by Western blot with anti-p62, ATG10, or β–actin antibodies. Differences between means were considered statistically significant when p<0.05 by Mann-Whitney U-test.