Fig 1. Reciprocal regulation of AnxA6 and GRF2 by non-selective calcium channel blockers.
A) BT-549 (AnxA6 high) and empty vector or AnxA6 expressing HCC1806 (AnxA6-low) cells were treated with Ni2+ or Ca2+ for 72 h and AnxA6, AnxA2 and GRF2 protein assessed by immunoblotting. B) Densitometric analysis of AnxA6 and GRF2 expression. Bars represent fold change relative to control cells from two independent experiments. C and D) Effects of Ni2+ concentration and time on the reciprocal expression of AnxA6 and GRF2. AnxA6 expressing HCC1806 cells were treated with the indicated concentrations of Ni2+ for 72 h (C) or at 0.5 mM Ni2+ for the indicated times (D); Densitometric analysis of the expression of AnxA6, GRF2 and AnxA2 from a representative experiment (C and D, left panels). E) HCC1806 cells were treated with the indicated concentrations of bepridil or amlodipine for 72 h and the expression of AnxA6 and GRF2 assessed by western blotting. F and G) Densitometric analysis of the protein bands in bepridil treated (F) and amlodipine treated cells from a representative experiment (G). Flag-AnxA6: flag-tagged recombinant AnxA6, End-AnxA6: endogenously expressed AnxA6.