Figure 5

IKK dependency of co-localized p65 and H3pSer10. (a) Confocal analysis of the extent of co-localization of p65 signal (green) with that corresponding to the phosphorylated Serine 10 of the histone H3 (red) in both basal conditions and after 60 min stimulation with 2 ng/ml IL-1β. The distinct signals are shown, together with the merged images including Hoechst 33342 as a nuclear counterstaining and a ×3 zoom in order to better appreciate the co-localized signal at the nuclear level, appearing as pink dots. (b) Analysis of the extent of co-localization derived from 26 different images for each condition, spread in the whole samples. Data are reported as mean ± SD. The means of the groups were compared by ANOVA, followed by Tukey’s post hoc test. The differences were considered significant when p < 0.05 with *p < 0.05; **p < 0.01; and ***p < 0.001. In both IKK KD, the IL-1β dependent induction is greatly blunted compared to the IKK proficient GL2 chondrocytes. Noteworthy, co-localized signals were significantly lower in IKKαKD compared to IKKβKD in both basal conditions and after IL-1β stimulation. IKKαKD data after IL-1β were significantly lower compared to both GL2 and IKKβKD samples.