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. 2015 Jun 11;10(6):e0129485. doi: 10.1371/journal.pone.0129485

Fig 6. Effect of IL-1β siRNA on nuclear DNaseI translocation in RPTEC stimulated with TNFα.

Fig 6

Confocal microscopy of sham-stimulated RPTEC (A, upper panels), cells stimulated with 20ng/ml of TNFα for 48 hrs (A, middle panels) and cells transfected with IL-1β specific siRNA and stimulated with TNFα for 48 hrs was performed by using anti-Trap1 antibodies (A, green) and by anti-DNaseI antibodies (A, red). While incubation of RPTEC with TNFα induced nuclear DNaseI translocation (A, middle panels), in RPTEC transfected with IL-1β siRNA and then stimulated with TNFα this did not result in translocation of DNaseI into the nuclei (A, lower panels). mRNA levels of DNaseI and IL-1β were increased in non-transfected RPTEC after stimulation with TNFα (B). Transfection with IL-1β siRNA inhibited mRNA level of IL-1β in RPTEC (C) but did not affect an increase in DNaseI gene expression induced by TNFα stimulation (C). Efficiency of the transfection procedure in RPTEC was controlled by transfection of P62 siRNA. As demonstrated in D, mRNA level of P62 was markedly decreased in transfected cells. Significances: *<0.05; **<0.005.