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. 2014 Sep 3;55(9):5551–5560. doi: 10.1167/iovs.14-14580

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Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

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The effect of DJ-1 downregulation on Nrf2-associated Cul3 and Keap1 levels. Immunoprecipitation–WB analysis of Cul3/Nrf2 and Keap/Nrf2 levels in non–siRNA-treated, control siRNA-treated, and DJ-1 siRNA-treated HCECi cells. (A) Left panel: IP-WB analysis of cell lysates immunoprecipitated with anti-Nrf2 antibody followed by immunoblotting with anti-Nrf2 antibody, anti-Cul3 antibody, and anti-Keap1 antibody. Middle panel: WB analysis of input immunoblotted with anti-Cul3 antibody, anti-Nrf2 antibody, anti-Keap1 antibody, and anti–DJ-1 antibody. Right panel: IP-WB analysis of non–siRNA-treated cells immunoprecipitated with anti-Nrf2 antibody (left lane) or normal rabbit IgG (IgG control, right lane) followed by immunoblotting with anti-Nrf2 antibody. Averaged densitometric analyses showed that DJ-1 decrease does not change Cul3/Nrf2 (B) and Keap1/Nrf2 (C) levels.