Figure 6

EMSA analysis of Sp1 binding activity in nuclear proteins of HAM. Panel A : HAM were treated or not with LPS (1 μg/ml) during 2 h. Nuclear proteins were extracted following procedure described in Materials and Methods. Lane 1: free labelled probe only (without nuclear proteins) – Lane 2: Nuclear proteins from control HAM incubated with labelled probe specific for Sp1 – Lane 3: idem lane2 except that HAM were treated 2 h with LPS – Lane 4: idem lane 3 except that an excess of unlabelled (cold) probe was added in the binding reaction – Lane 5: idem lane 3 except that a mutant labelled probe (mutation in consensus binding site) is used instead of the specific labelled probe for Sp1 – Lane 6: idem lane 3 except that nuclear proteins were preincubated with a specific anti-Sp1 antibody before addition of the specific labelled probe. Panel B represents a quantitative analysis of a EMSA gel from HAM following stimulation by LPS. Data are mean ± SD from 6 experiments; *p value = 0.0411 (Mann-Whitney U test).