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.