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. 2000 Jan;99(1):54–61. doi: 10.1046/j.1365-2567.2000.00943.x

Figure 2.

Figure 2

Binding profiles of CD154/wt and CD154/G219R proteins to different reagents. (a). 293 cells were transiently transfected with 20 µg CD154/wt or CD154/G219R plus 5 µg pMCD8. Equal numbers of cells were analysed 48 hr later for expression of CD154 by staining with CD40‐Ig (panel 1), anti‐TRAP mAb (panel 2), and a rabbit polyclonal anti‐TRAP antiserum (panel 3). Cells were also stained with FITC‐labelled OKT8 to control for transfection efficiency (panel 4). Dotted peaks in each panel represent background staining with either isotype control mAb (panels 2 and 4) or secondary antibodies alone (panels 1 and 3). (b). Data was analysed from multiple (> 3) independent transfections to establish the level of recognition of the CD154/G219R protein by the different reagents. The transfection efficiency within an experiment was normalized to the expression of CD8. Error bars represent SEM. Differences in recognition of CD154/G219R and CD154/wt by CD40‐Ig, mAb anti‐TRAP, and mAb 24‐31, were determined to be statistically significant to a 95% confidence level by a standard paired t‐test (P < 0·02). The difference in binding for polyclonal anti‐TRAP and mAb 5c8 to either CD154/G219R or CD154/wt was determined not to be significant (P < 0·8 and P < 0·33, respectively).