Figure 4

In vitro interaction assay of IN mutants with LEDGF/p75 and TNPO3. Interaction between INs and LEDGF/p75 (A) or TNPO3 (B) was measured by HTRF. GST-LEDGF/p75 (5 nM) or GST-TNPO3 (5 nM) and His-IN WT or indicated IN mutants (80 nM) were incubated with anti-GST-Europium Cryptate and anti-His-XL antibodies. HTRF signal was read at 2 h post incubation in a Pherastar (BMG) at 665 and 620 nm after excitation at 337 nm. HTRF signals of indicated IN mutants were expressed as a percent IN WT ΔF. Results are an average of three (IN::LEDGF/p75) or four (IN::TNPO3) experiments performed in duplicate. Statistical significance values were calculated using a student two-sided t test. ***, P < 0.0001; **, P < 0.001; *, P < 0.01.