Fig. 4

p68 and β-catenin synergism potentiates NF-κB signaling axis. a HCT 116 (upper panel) and SW-480 (lower panel) cells were seeded in 100 mm plates and transfected with 8 μg of EV or pGZ-p68. Cytoplasmic and nuclear extracts were prepared, 36 h post transfection; and probed for the indicated proteins by IB. Densitometry values relative to loading controls (α-tubulin for cytoplasmic and Lamin B for nuclear extract, respectively) are given below the blots. b Multiple colon cancer cell lines, grown in 60 mm cell culture dishes were transfected with 4 μg of pGZ-p68 or EV. ‘Exo’ and ‘Endo’ depicts exogenous and endogenous p68, respectively. c Colon cancer cell lines, as indicated were seeded in 60 mm cell culture dishes and transfected with 4 μg of scr or p68 shRNA1 and p68 shRNA2 constructs. d HCT 116 cells were seeded in 60 mm plates and were transfected with either scr or p68 siRNA in a dose-dependent manner as indicated. e Colon cancer cell lines, grown in 60 mm cell culture dishes were transfected with 4 μg of either pGZ-β-catenin or EV. ‘Exo’ and ‘Endo’ depicts exogenous and endogenous β-catenin, respectively. f Colon cancer cell lines, grown in 60 mm plates were transfected with 4 μg of either scr or β-catenin shRNA construct. g HCT 116 cells were seeded in 60 mm plates and were transfected with either scr or TCF4 siRNA in a dose-dependent manner as indicated. h HCT 116 cells were seeded in 60 mm plates and were transfected with 2 μg of either scr or p68 siRNA, in combination with 2 μg of pGZ- β-catenin or EV. Cell lysates were prepared from 36 h (b & e) and 48 h (c, d, f, g & h) post transfected cells and probed for the indicated proteins by IB as depicted in different sub-figures. Densitometry values below the blots in panels (b-h) are relative to the loading control (Actin). In sub-figures (d & g) - 2 and 4 μl siRNAs were used from 10 μM stock