FIGURE 6.

Fumarate mediates HIF-1α mRNA expression through TBK1. A, FH or fumarate hydratase + Tank-binding kinase-1 (FH + TBK) were transiently silenced (siFH and siTBK/siFH, respectively) in DKO (IKKα and IKKβ) MEFs. Scrambled oligos were transfected as a negative control. After 48 h, cell lysates were prepared and analyzed for phospho-p65 Ser⁵³⁶ (p-p65 S⁵³⁶), total p65, total TBK and FH. Tubulin was used as loading control. B, HK2 cells were treated with DEF simultaneously with the Tank-binding kinase inhibitor BX795 at the indicated concentrations for 8 h. Total protein was used to assess HIF-1α, phospho-p65 Ser⁵³⁶ expression. Tubulin was used as loading control.