Figure 1.

PAF or IL-1β differentially enhance DNA binding activities of cis-acting transcription factors. EMSA using NPXTs from HEK cells reacted with target DNA oligonucleotide consensus or with mutant sequences for the transcription factors AP1, AP2, Egr1 (zif268), GAS, NF-κB, SIE, and TFIID described in Table 1. Leftmost lane represents the migration of a typical free ³²P end-labeled oligonucleotide with no NPXT. Relative gel shift signal quantitation in the bar graph was for gel-shifted species unique to that transcription factor-DNA binding consensus sequence, minus the signal contributed by the mutant DNA, and located in between the two sets of arrows. NF-κB quantitation includes only the upper (p50-p65) activator complexes whose identity was confirmed by using p50 and p65 antibodies and gel supershift assay (data not shown). Gel is overexposed to show the relative signal intensities among each of the shifted species. Exposure time = 12 hr. n = 4; mean ± SD; significance of induced factors over control, ∗, P < 0.001; significance of AP2, Egr1, and SIE over control P > 0.05; significance of TFIID over control, P > 0.68, ANOVA.