Objective: To demonstrate that CaMKK mediates LPSinduced TNFα production through interactions with the mitogenactivated protein kinase(MAPK) cascade Methods: RAW 264 cells were stimulated with LPS(10 ng/mL). CaMKK, CaMK I and IV were inhibited with either KN62(all CaMK), STO609(CaMKK) or with targeted siRNA. TNFα was assayed by ELISA. MAPK activation and protein knockdown was assessed by western blot. Statistical analysis was performed by ANOVA. Results: LPS-induced TNFα production was inhibited by 85% by either KN62 or STO609. CaMKK and CaMK IV siRNA inhibited cytokine production, which was significant for CaMKKb. All MAPK were activated by LPS, but only JNK phosphorylation was abrogated with STO609 inhibition of CaMKK. Conclusion: CaMKK mediates LPS-induced TNFα production, most likely through the CaMKK cascade of CaMK IV. This may also involve interactions with JNK of the MAPK family.