Involvement of P53 and Bax/Bad triggering apoptosis in thioacetamide-induced hepatic epithelial cells

Figure 1 Cell viability of clone 9 after treatment with 100 mg/L TAA for various periods. The results were obtained using MTT assay. Data are presented as mean ± SE. Individual experiment was repeated three times and each time point of treatment was triplicate.

Figure 2 Total glutathione (GSH and GSSG) of clone 9 after treatment with 100 mg/L TAA for various times. Data are presented as mean ± SE. ^bP < 0.001 vs the control (0 h). Individual experiments were repeated three times and each time point of treatment was triplicated.

Figure 3 A: Caspase 3 activity; B: Caspase protein level (Western blot); C: Alterations of caspase protein compared to H0 (zero hour) after β-actin normalization (Western blot) in clone 9 after treatment with 100 mg/L TAA for various times. Data are presented as mean ± SE. ^aP < 0.05 vs the control (0 h); ^bP < 0.01 vs the control (0 h). Individual experiments were repeated three times and each time point of treatment was triplicate.

Figure 4 A: Morphology of clone 9 cell; (a) untreated (Hoechst 33342 staining); (b) after treatment with 100 mg/L TAA treatment for 16 h (light field); (c) after treatment with 100 mg/L mmol/L TAA for 16 h (Hoechst 33342, 5 mg/mL in PBS). The arrow indicates the apoptotic cells; B: Flow cytometry of apoptosis in clone 9 cells (TUNEL assay) after treatment with 100 mg/L TAA for various times (a) 0 (b) 4 and (c) 8 h. R1 presents the area of cell apoptosis. Individual experiments were repeated three times and each time point of treatment was triplicate.

Figure 5 A: Phospho-p53 protein level (typical data, Western blot); B: Alterations of phospho-p53 protein as compared to H0 following β-actin normalization of clone 9 after treatment with 100 mg/L TAA for various times. Data are presented as mean ± SE. ^aP < 0.05 vs the control (0 h). Individual experiments were repeated three times and each time point of treatment was triplicate.

Figure 6 A: Bax protein level (typical data, Western blot); B: Alterations of Bax protein as compared to H0 following β-actin normalization of clone 9 after treatment for various times with 100 mg/L TAA. Data are presented as mean ± SE. ^aP < 0.05 vs the control (0 h). Individual experiment was repeated three times and each time point of treatment was triplicate.

Figure 7 A: Bad protein level (typical data, Western blot); B: Alterations of Bad protein as compared to H0 following β-actin normalization of clone 9 after treatment for various times with 100 mg/L TAA. Data are presented as mean ± SE. ^aP < 0.05 vs the control (0 h). Individual experiments were repeated three times and each time point of treatment was triplicate.