Figure 3

Apoptotic cell death and HO-1 expression in CC cells treated with PTL and/or Ro. (A) Apoptotic cell death in Choi-CK and SCK cells treated with PTL and/or 5 µM Ro for 72 h. The cells were stained with Hoechst 33258, and the fraction of apoptotic cells was determined using fluorescence microscopy. Each data point represents the mean ± SE of three independent experiments performed in duplicate. ^*, P < 0.05; ^**, P < 0.01. (B) Quantification of the apoptotic fraction by FCM analysis. Cells were treated with PTL and/or 5 µM Ro for 72 h. The sub G₁ fraction was estimated by gating hypodiploid cells in the histogram using the LYSIS II program. DNA content is plotted on the linear abscissa (M₁, apoptotic fraction). Each value represents the mean ± SE of three independent experiments performed in duplicate. (C) Expression of Nrf2 and HO-1 in Choi-CK cells treated with PTL and/or 5 µM Ro for 72 h. (D) Expression of Nrf2 and HO-1 in SCK cells treated with PTL and/or 5 µM Ro for 72 h. Immunoblot experiments were performed at least three times, and the result of one representative experiment is shown.