Increased DNA binding activity of NF-kB, STAT-3, SMAD3 and AP-1 in acutely damaged liver

Figure 1 Semiquantitative RT-PCR analysis for iNOS expression. A: PCR products analyzed by agarose electrophoresis; B: normalized values of iNOS expression with respect to the housekeeping gene HPRT in all analyzed groups of three different PCR reactions performed.

Figure 2 Semiquantitative RT-PCR analysis for HGF and c-met expressions. A: PCR products analyzed by agarose electrophoresis; B: Normalized values of HGF and c-met expression with respect to the housekeeping gene HPRT in all analyzed groups of three different PCR reactions performed.

Figure 3 Binding activity of NF-κB in acute liver damage. Wistar rats were intoxicated acutely with CCl₄ and then sacrificed at different time points. Nuclear proteins were extracted from the livers and Gel-shift assay was performed for NF-κB transcription factor. A: Binding DNA activity of NF-κB after CCl₄ intoxication; B: supershift assay using 1-h sample, 1 μL of polyclonal antibody against p65 subunit was added and incubated for 1 h before usual EMSA assay; C: densitometric analysis of results of EMSA assay.

Figure 4 Binding activity of STAT-3 in acute liver damage. Wistar rats were intoxicated acutely with CCl₄ and then sacrificed at different time points. Nuclear proteins were extracted from the livers and Gel-shift assay was performed for STAT-3. A: Binding DNA activity of STAT-3 after CCl₄ intoxication; B: supershift assay similar using 6-h sample; C: densitometric analysis of results of EMSA assay.

Figure 5 Binding activity of SMAD3 in acute liver damage. Gel-shift and supershift assays were performed to analyze SMAD3 DNA binding activity. A: Binding DNA activity of SMAD3 after CCl₄ intoxication; B: supershift assay using 24-h sample; C: densitometric analysis of results of EMSA assay.

Figure 6 Binding activity of AP-1 in acute liver damage. EMSA and supershift assays, similar to Figures 3 and 4, were performed to analyze AP-1 activity in the same samples. A: Binding DNA activity of AP-1 after CCl₄ intoxication; B: supershift assay using 24-h sample; C: densitometric analysis of results of EMSA assay.