FIG 5.

US27 promotes ARE-driven transcription. (A) HEK293 or 293-US27 stable cell lines were transfected with the pCignal-ARE-GFP plasmid, and the relative fluorescence intensity (RFI) was measured by flow cytometry. (B) HEK293 and stable cell lines were transfected as described above, and the relative fluorescence intensity was measured after 48 h. *, P < 0.01 by paired Student's t test. (C to F) HEK293 and 293-US27 cells were transfected with the pCignal-ARE-GFP plasmid and after 24 h were transfected with the indicated siRNAs for 48 h. (C) RNA was extracted, RT-PCR was performed with the indicated primers, and the products were visualized via agarose gel electrophoresis. (D) Immunoblotting of cell lysates with the indicated siRNA treatments. (E) The relative fluorescence intensity was measured by flow cytometry. *, P < 0.01 by paired Student's t test versus all antibodies; a, significance versus NRF-1; b, significance versus NRF-2; c, significance versus NRF-1/2. (F) Data from panel E are expressed as percent total fluorescence. *, P < 0.01 by paired Student's t test. Error bars represent the standard error for three replicates.