Overexpression of Hsc70 increases nuclear p65 and elevates the activity of NF-κB.

<p><b>A.</b> HEK293FT cells were transfected with expression vectors as indicated. An inactive state of NF-κB (baseline) was achieved by co-transfection of IκB expression vector. The nuclei and the whole cell body were defined as regions and their mean fluorescence was measured using ImageJ-software. Co-expression of IκB significantly reduces nuclear Hsc70 GFP, whereas no effect on the nuclear localization of GFP was observed. <b>B.</b> HEK293FT cells were transfected as indicated in A. Co-expression with Hsc70 GFP significantly increased nuclear p65-FPred compared to co-expression with GFP. No increase of nuclear FPred p65 was observed if additional IκB was expressed (data not shown). <b>C.</b> Luciferase reporter assay revealed a significant increase of NF-κB activity in HEK293 cells co-transfected with Hsc70 and p65 (+32%), compared to cells over-expressing p65 alone.