Induction of apoptosis and expression of Bcl-2 in MS4A-IκBα32/36A-transfected cells. (A) Analysis of nuclear condensation after Hoechst 33258 staining. Cells were (+ DoxyC) or were not (− DoxyC) pretreated with doxycycline at 2 μg/mL for 24 hours. Daunorubicine (Dauno) was added for 18 hours at the concentrations indicated. (B) Dose response to apoptosis induction by daunorubicine in cells pretreated (+ DoxyC) or not (− DoxyC) with doxycycline at 2 μg/mL for 24 hours. Percentages of apoptotic cells were assessed by counting cells with apoptotic nuclei under fluorescence microscope after Hoechst 33258 staining. (C) Flow cytometry analysis of DNA content after propidium iodide staining of the DNA of cells pretreated (+ DoxyC) or not (− DoxyC) with doxycycline at 2 μg/mL for 24 hours and then treated with daunorubicine at 0.2 μmol/L for 18 hours. (D) Annexin V staining of cells pretreated (+ DoxyC) or not (− DoxyC) with doxycycline at 2 μg/mL for 24 hours and then treated with daunorubicine at 0.2 μmol/L for 18 hours. (E) Bcl-2 expression of cells treated (+) or not (−) with doxycycline at 2 μg/mL for 48 hours was assessed by Western blot.
