Fig. 5

Cytokines upregulate GFPT2 mRNA expression in an NF-κB-dependent manner. a & b A549 spheroid cultures upregulate GFPT2 mRNA expression and protein expression in response to TNF, or combination of TNF and TGF. c & d A549 cells transduced with adenovirus encoding a non-degradable form of the NF-κB inhibitor, SR-IκBα (+) were unable to upregulate GFPT2 mRNA expression in response to TNF or TNF/TGF, relative to GFP control (−) cells. e & f A549 cells treated with Bay 11–7085 (5 μM) or vehicle (DMSO) for one hour, prior to a two hour stimulation with TNF/TGF, show that IKK activity was required to increased GFPT2 mRNA and protein levels. f Detection of total IκBα protein levels was used as a measurement of IKK activity. RT-qPCR analysis represents three independent experiments; mRNA levels were calculated relative to HPRT mRNA expression. RT-qPCR performed on GFPT1 and TNFAIP3 served as negative and positive NF-κB regulated gene targets, respectively. Mean + SD are shown * p = < 0.05, and ** = < 0.01; ns = not significant relative to controls. Immunoblots were analyzed for protein loading controls, GAPDH or Tubulin