<p>Assays using site directed mutagenesis of AP-1 and NFκB binding sites. (<b>A–I</b>) Schematic of each mutation of CR1 construct. Mutated sites are identified by a red X. (<b>A’–I’</b>) Transfection of each the constructs in SUM159 cells. (<b>J</b>) Quantification of the number of GFP-expressing cells/total number of cells counted. Control mutation at a non-conserved site (<b>B’</b>) showed no difference in GFP expression when compared to CR1 (<b>A’</b>). Single site mutations of AP-1-1, AP-1-2 and NFκB (<b>C’-E’</b>) showed a significant reduction of GFP expression compared to CR1. However, GFP expression was not eliminated entirely. Mutation of a combination of AP-1 and NFκB binding sites (<b>F’-H’</b>) did not reduce further GFP expression, however, the percentage of GFP expression was still significantly reduced compared to CR1. Mutation of all three TFBSs (<b>I’</b>) showed the greatest reduction of GFP expression. **p = < 0.0005 ***p = <1.0×10<sup>−5</sup> (student’s t-test). Scale bar = 50 µM.</p
