Figure 1
From: ERK2 phosphorylation of serine 77 regulates Bmf pro-apoptotic activity
Bmf induces apoptosis in melanoma cells independent of Bim-EL. (a) WM793TR/HA-Bmf, Sbcl2TR/HA-Bmf, 1205LuTR/HA-Bmf, WM115TR/HA-Bmf and A375TR/HA-Bmf cells were treated with or without 100 ng/ml doxycycline for 7 h. Cell lysates were analyzed by western blotting using HA-tag and actin (loading control) antibodies. (b) As for a, except that cells were induced with or without 100 ng/ml doxycycline for 48 h. Cells were then stained with annexin-V-APC for flow cytometry analysis. Quantitated are the mean percentages of annexin-V-positive cells from three independent experiments. Error bars: standard deviation. (c) WM793TR/HA-Bmf cells were transfected with control or Bim siRNA for 72 h. Cells were then treated with or without 100 ng/ml doxycycline for additional 24 h before lysis for western blot analysis. (d) As in c except that after 24 h of doxycycline treatment, cells were subject to annexin-V staining and flow cytometry analysis. Representative flow traces from two independent experiments flow traces are shown. x axis, fluorescence intensity; y axis, cell numbers
