To further investigate the regulation role of two chemokine genes CCL3 and CCL4 in chondrocytes in response to resistin, human primary chondrocytes and T/C-28a2 cells were cultured. The function of resistin on the chemokine genes, and the expression of C/EBP beta, NF-kappa B isoforms were tested using qPCR. The methods used to investigate timed co-regulation of C/EBP beta and NF-kappa B were NF-kappa B inhibitor (IKK-NBD) and C/EBP beta inhibitor (SB303580) treatments, and subcellular localization, with or without resistin stimulation. Results showed that resistin could increase the up-regulation of chemokine genes independently. Resistin increased the expression of C/EBP beta and NF-kappa B isoforms. C/EBP beta regulated basal activity and steadily increased over time up to 24h with resistin. NF-kappa B was up-regulated upon induction with resistin, peaking at 4 h. C/EBP beta and NF-kappa B co-enhanced the chemokines expression; inhibition of their activity was additive. The timing of activation in chondrocytes was confirmed by subcellular localization of C/EBP beta and c-rel. Chondrocytes react to resistin in a non-restricted cell-specific manner, utilizing C/EBP beta and NF-kappa B in a combinatorial regulation of chemokine gene expression. The activity of C/EBP beta is augmented by a transient increase in activity of NF-kappa B, and both transcription factors act independently on the chemokine genes, CCL3 and CCL4. Thus, resistin stimulates CCL3 and CCL4 through combinatorial regulation of C/EBP beta and NF-kappa B in chondrocytes.