KLF3 from erythroid and non-erythroid cells display similar DNA-binding abilities in vitro. (A) EMSAs were employed to assess the binding of KLF3 to the murine α-globin promoter (lanes 1–7) and a site in the HS-26 element from Figure 5 (lanes 8–14). Nuclear extracts were harvested from non-erythroid MEF (lanes 4, 5, 11 and 12) or erythroid MEL cells (lanes 6, 7, 13 and 14). Nuclear extracts from mock transfected COS-7 cells (lanes 1 and 8) or cells expressing KLF3 (lanes 2, 3, 9 and 10) were included as negative and positive controls respectively. The identity of KLF3 was confirmed by specific antibody supershifts (lanes 3, 5, 7, 10, 12 and 14). (B) Western blot demonstrating the relative amounts of KLF3 in MEF (lane 4) and MEL (lane 5) nuclear extracts used in the EMSAs in (A). As negative and positive controls, COS and COS-KLF3 nuclear extracts have been included (lanes 2 and 3) at 20-fold lower relative amounts than in (A) to facilitate visualization. A size ladder is shown in lane 1.
Funnell et al. BMC Molecular Biology 2014 15:8 doi:10.1186/1471-2199-15-8