Figure 1

IKK dependency of the transcription of monocyte recruiting chemokines. (a) Total RNA obtained from GL2, IKKαKD or IKKβKD chondrocytes in basal conditions or following 8 h exposure to 2 ng/ml IL-1β was probed against the Oligo GEArray microarray human chemokines and receptors microarray (OHS-022, SuperArray). The array layout is indicated in Supplementary Fig. 2. The probe positions for the monocyte active chemokines (CCL2/MCP-1, CCL3/MIP-1α, CCL4/MIP-1β and CCL5/RANTES) are indicated by red arrows. (b) For each chemokine, every single dot signal of the quadruplicate was quantified by densitometric measurements, and signals were compared across the different conditions: GL2 (black pattern), IKKαKD (gray pattern) or IKKβKD (white pattern) chondrocytes in control (−) or IL-1β stimulation (8 h, +). The different Y axis scale shows the different expression magnitudes of these chemokines in the three different genotypes. Cumulative results of the four replicates are shown as mean ± SD. Chemokine gene expression levels in both basal and IL-1β stimulated conditions were compared across the different phenotypes by ANOVA, followed by Tukey’s post hoc test. The differences were considered significant when p < 0.05 with: **p < 0.01; and ***p < 0.001. (c) Real Time PCR performed with RNA obtained from cultures derived from three different patients confirms an evident IL-1β induction and in basal conditions lower CCL2/MCP-1 RNA in IKKαKD compared to the GL2 controls.