siRNA unrelated AdipoR1 AdipoR2 None Ad Osm A9 None Ad Osm A9 None Ad Osm A9 Ratio ** Protein (μM) None Ad Osm A siRNA Unrelated AdipoR1 AdipoR2 Osmotin Induces AMPK Activation In Murine C2C12 Myocytes Via Adiponectin Receptors Narasimhan et al, Mol Cell (2005) 17: Ad AMPK-P AMPK OSM Osmotin Mimics Adiponectin the muscle

Improved mitochondrial function Increased mitochondrial number Increased expression PGC1α Increase ATP production Increased fatty acid oxidation Increased glucose catabolism Reduction of Blood glucose and fatty acids Antidiabetic Adiponectin AMPK-P↑ Adiponectin receptor the muscle Adiponectin Improves Mitochondrial Function In Human Muscle Cells (Antidiabetic) A. E. Civitarese

Osmotin induces Beta Oxidation activity (3h) control1ug/ml5ug/ml βHAD Activity (umole/min/mg protein) Osmotin Promotes Fatty Acid Oxidation In Human Biopsy Muscle Cells Fatty acid Acetyl CoA TCA cycle ATP βHAD A. E. Civitarese

Osmotin induces TCA Cycle activity control1ug/ml2.5ug/ml Citrate Synthase Activity (umole/min/mg protein) Osmotin increases mitochondrial number Control1ug/ml2.5ug/ml mtDNA/nuclear genome Osmotin Promotes Glucose Catabolism in Human Biopsy Muscle Cells Glucose mitochondria TCA cycle A. E. Civitarese

Osmotin Promotes AMPK Activation And PGC1α Gene Expression in Human Biopsy Muscle Cells AMPK-P Total AMPK PGC1α β-actin Osmotin (µg/ml) A. E. Civitarese

Improved mitochondrial function Increased mitochondrial number Increased expression PGC1α Increase ATP production Increased lipid oxidation Increased glucose uptake Reduction of Blood glucose and fatty acids Antidiabetic Osmotin AMPK-P↑ Adiponectin receptor Osmotin Improves Mitochondrial Function In Human Muscle Cells (Antidiabetic) Osmotin Mimics Adiponectin the muscle

Osmotin Improved Hyperglycemia In db/db Mice Relative Plasma Glucose Levels min Test materials delivered by intraperitoneal injection PBS Osmotin Yamauchi et al., 2001, Nat Med 7: Yamauchi et al, unpublished data Osmotin Mimics Adiponectin

Crohn’s disease (colitis, usually of small intestine), Diarrhea, Abdominal pain Weight loss, Anemia, Inflammation Stomach Colon Small intestine Rectum Plasma Adiponectin Levels Have Inverse Correlation With The Severity Of Colitis Plasma Adiponectin ↓ Severity of Colitis ↑ ↑↓

Body Weight Gain DSS+ SAL DSS+ OSM % body weight gain Water DSS+ OSM DSS+ SAL Water Percent Hematocrit Osmotin Treatment Reduces Weight Loss And Anemia Associated With Induced Colitis In Mice colitis * * R. Arsenescu

Plasma IL-1β (Pro-inflammatory cytokine) DSS+ OSM DSS+ SAL WATER pg/ml Osmotin Treatment Reduces Inflammation Associated With Induced Colitis In Mice DSS+ OSM DSS+ SAL WATER pg/ml Plasma IL-10 (Anti-inflammatory cytokine) colitis * R. Arsenescu