Figure 4: Modulation of LPS activated signaling pathways by NT at 10 (a) and 30 mM glucose (b), in macrophages, by western blot (A) and relative quantification (B). Cells were plated at 1.5 × 106/well and treated simultaneously with 10 nM NT and 1 μg/mL LPS during 5, 15, 30, or 60 minutes. The lysates were probed for phospho p38 MAPK, phospho p44/42 MAPK, phospho pAKT (Ser 437), and inhibitory protein for NF-κB activation, IκB-α antibodies. Equal amounts of protein were evaluated with total p38 MAPK, p44/42 MAPK, AKT, and actin antibodies. The results shown are representative of four to six independent experiments with similar results.