Figure 5
From: Maternal smoking and the retinoid pathway in the developing lung
Cigarette smoke condensate (CSC) decreases retinoic acid response element binding in vitro. A: Representative electrophoretic mobility shift assay (EMSA) showing decreased binding of nuclear extracts from A549 cells to the DR5 retinoic acid response element after 1 μM retinoic acid and vehicle (lanes 2 – 5) or 0.50% cigarette smoke condensate (CSC, lanes 6–9) treatment followed by second CSC dose of 0.50% 14 h after the first, with extracts harvested at 15 – 120 min. following this second dose. Similar results were also shown in an independent replication. B: Relative densities of A549 cell nuclear extract/DR5 retinoic acid response element complexes (normalized to free probe) following stimulation with 1 μM retinoic acid and either vehicle or 0.5% CSC as outlined for panel A. Data are mean ± SEM from two independent experiments. Abbreviations: N = negative control lane (lane 1) with equivolume extraction buffer substituted for the nuclear extract; numbers indicate time course in minutes, lane numbers for samples indicated below figure, NE = nuclear extract, CSC = cigarette smoke condensate.