Fig 2: Schematic illustration of CBP bromodomain inhibitor prevents sepsis-related HMGB1 secretion and blocks HMGB1 pro-inflammatory activity. The potent and selective CBP inhibitor, SGC-CBP30, significantly rescues mice from LPS-/CLP-induced sepsis model. In addition, SGC-CBP30 suppresses LPS-induced HMGB1 expression and cytoplasmic translocation. Furthermore, by triggering the reactivation of PP2A and the stabilization of MKP-1, CBP bromodomain inhibition prevents rhHMGB1-stimulated activation of MAPKs and NF-κB pathways and production of proinflammatory cytokines.

Fig 3: UT extract attenuates FFA induced hyperactivity of PP2A and enhances Akt phosphorylation in C2C12 myotubes.Fully differentiated C2C12 myotubes were incubated with palmitate (250 uM) with or without UT (5 ug/ml) or 10 nM okadaic acid (OK) for 16 hours. (a) Cells were extracted for lipids by the Folch partition and ceramide quantities determined by tandemn mass spectrometry (LC/MS/MS). (b) Cell lysates were used to determine PP2A activity by a standard kit (EMD Millipore, Temecula CA, USA). (c) Cell lysates were separated by SDS-PAGE and analyzed by immune blot analysis for PP2A-A and PP2A-B protein expression and (d) PP2A-C protein expression, PP2AC tyrosine phosphorylation and PP2A-C leucine methylation. Each panel represents an experiment independently repeated three times on different batches of myotubes. *denotes significant difference with FFA only treatment (P < 0.05). #denotes significant difference with the untreated control treatment (P < 0.05).

Fig 4: Changes in the renal Gnmt expression and SAM content. Representative western blots of Gnmt and β-actin in the renal cortex (A). Quantitative ratios of Gnmt to β-actin (B) and mRNA expression of Gnmt adjusted to 18S levels (C) in the renal cortex (n=7). Representative photographs of SAM immunohistochemistry of the kidney (scale bar: 100 μm) (D). Semiquantitation of the SAM staining scores (E) (n=3). SAM levels in the renal cortex (F) (n=3). Representative western blots of LCMT1, methylated-PP2A, PP2A and β-actin in the renal cortex (G). Quantitative ratios of LCMT1 to β-actin (H) and methylated-PP2A to PP2A (I) in the renal cortex (n=7). The data shown are the means ± SD. **p<0.01, ***p<0.001 vs. the indicated groups. ns: not significant. Gnmt: glycine-N-methyltransferase, SAM: S-adenosylmethionine, LCMT1: leucine carboxyl methyltransferase 1, PP2A: protein phosphatase 2A.

Fig 5: UT extract attenuates HFD induced hyperactivity of PP2A and enhances Akt phosphorylation in skeletal muscle.Male C57BL/6J mice were fed were fed the experimental diets for 12 weeks. After sacrifice (a) 30 mg of gastrocnemius muscle was extracted for lipids by the Folch partition and ceramide quantities determined by tandemn mass spectrometry (LC/MS/MS). (b) 30 mg muscle was homogenized and lysates used to determine phosphatase PP2A activity by a standard kit (EMD Millipore, Temecula CA). (c) Protein homogenates from 20 mg gastrocnemius muscle were separated by SDS-PAGE and analyzed by immune blot analysis. Representative blots for PP2A- C protein expression, PP2A-C tyrosine phosphorylation and PP2A-C leucine methylation are shown. Fold change relative to LFD alone for each protein was calculated and mean ± SEM graphed (n = 11). #denotes significant difference with HFD (P < 0.05).