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. 2005 Jan;25(1):162–171. doi: 10.1128/MCB.25.1.162-171.2005

FIG. 7.

FIG. 7.

NRF2 accumulated when Cul3 or Keap1 was silenced. (A) 293T cells were cotransfected with the plasmid expressing FLAG-tagged NRF2 and the indicated short hairpin RNA plasmids (shCul3 and shKeap1). Transfected cells were selected by puromycin (1 μg/ml). Ninety-six hours after transfection, selected cells were lysed, and 50-μg samples of total lysates were resolved by SDS-PAGE and immunoblotted with the indicated antibodies. RNAi, RNA interference. (B) The expression vector for GFP-NRF2 alone or for GFP-NRF2 and DsRed-Keap1 was cotransfected with CUL3 short hairpin RNA plasmid or control short hairpin RNA plasmid to HeLa cells. After puromycin selection, the expression and localization of GFP-NRF2 and DsRed-KEAP1 were examined by fluorescence or phase-contrast microscopy.