Figure 2

Antioxidant treatment prevents LPS induced SIRT6 sulfenylation and glycolysis in LPS sensitive THP-1 cells, whereas prooxidant treatment enhances LPS induced SIRT6 sulfenylation and glycolysis in LPS tolerant THP-1 cells: (A–E), Effect of Febuxostat (FBX) treatment on Glycolysis in sensitive cells: (A) Cells were pretreated 30 minutes with 30 µM FBX followed by 3 hour LPS stimulation. Lysates were made and evaluated for SIRT6 sulfenylation (n = 3), (B) Cells were pretreated 30 minutes with 30 µM FBX followed by 6 hour LPS stimulation. Media was collected and evaluated for lactate production (n = 6), (C) Cells were pretreated 30 minutes with 30 µM FBX followed by 4.5 hour LPS stimulation. Membrane fractions were made and evaluated for Glut-1 expression (n = 3). (D,E) Cells were pretreated 30 minutes with 30 µM FBX followed by 6 hour LPS stimulation. Cells were evaluated for Glycolysis by Seahorse Respirometry (n = 3) (F–J), Effect of Tert-butyl hydroperoxide (TBH) treatment on Glycolysis in tolerant cells: (F) Tolerant cells were pretreated 30 minutes with 120 µM TBH followed by 3 hour LPS stimulation. Lysates were made and evaluated for SIRT6 sulfenylation (n = 3), (G) Tolerant cells were pretreated 30 minutes with 120 µM TBH followed by 6 hour LPS stimulation. Media was collected and evaluated for lactate production (n = 4), (H) Tolerant cells were pretreated 30 minutes with 120 µM TBH followed by 4.5 hour LPS stimulation. Membrane fractions were made and evaluated for Glut-1 expression (n = 3). *P < 0.05 (two-tailed Student’s t-test for pairwise comparisons), (I–J) tolerant cells were pretreated 30 minutes with 120 µM TBH followed by 6 hour LPS stimulation. Cells were evaluated for glycolysis by Seahorse Respirometry (n = 3).