Research Paper Volume 13, Issue 14 pp 17978—18005

Figure 5. IL-6 and IL-8 expression observed after SAHA treatment is partly dependent on NF-κB activation. Cells at early passage were treated with 0, 5 or 10 μM of SAHA during 24, 48 or 72 hr in combination or not with JSH-23 treatment (NF-κB inhibitor) during 24 hr. (A) Representative Western blots showing IĸBα total protein abundance after SAHA treatment. AG04431 dermal fibroblasts treated with 20 ng/mL of TNF-α during 20 minutes were used as positive control for IĸBα degradation. α-tubulin was used as loading control. (B) Quantification of the relative protein abundance of IĸBα. Signal intensities were quantified and normalized relative to the abundance of α-tubulin and are expressed relative to the control condition (0 μM SAHA, 24 hr). (C) Immunofluorescence analysis of p65 (green) nuclear translocation. Nuclei were labelled with DAPI (blue). Cells were visualized with confocal microscopy (scale bar = 50 μM). (D) Quantification of the percentage of p65 positive nuclei. (E) Steady-state mRNA level of IL-6 and IL-8. GAPDH was used as housekeeping gene. Results are expressed as fold induction in comparison with control fibroblasts (0 μM SAHA, 24 hr). Statistical analyses were performed using an ANOVA II (*: p<0.05; **: p<0.01; ***: p<0.001).