1. siRNA unrelated AdipoR1 AdipoR2 None Ad Osm A9 None Ad Osm A9 None Ad Osm A9 Ratio 32103210 ** Protein (μM) None Ad Osm A9 0.10 0.04 0.50 siRNA Unrelated AdipoR1 AdipoR2 Osmotin Induces AMPK Activation In Murine C2C12 Myocytes Via Adiponectin Receptors Narasimhan et al, Mol Cell (2005) 17: 171-180 Ad AMPK-P AMPK OSM Osmotin Mimics Adiponectin the muscle

2. 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

3. Osmotin induces Beta Oxidation activity (3h) 0.80 1.20 1.60 2.00 2.40 2.80 3.20 3.60 4.00 4.40 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

4. Osmotin induces TCA Cycle activity 0.80 1.20 1.60 2.00 control1ug/ml2.5ug/ml Citrate Synthase Activity (umole/min/mg protein) Osmotin increases mitochondrial number 0 50 100 150 200 250 Control1ug/ml2.5ug/ml mtDNA/nuclear genome Osmotin Promotes Glucose Catabolism in Human Biopsy Muscle Cells Glucose mitochondria TCA cycle A. E. Civitarese

5. Osmotin Promotes AMPK Activation And PGC1α Gene Expression in Human Biopsy Muscle Cells AMPK-P Total AMPK PGC1α β-actin Osmotin (µg/ml) 0 1 5 7.5 10 20 A. E. Civitarese

6. 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

7. Osmotin Improved Hyperglycemia In db/db Mice Relative Plasma Glucose Levels 120 100 80 60 0153060120 min Test materials delivered by intraperitoneal injection PBS Osmotin Yamauchi et al., 2001, Nat Med 7:941-946 Yamauchi et al, unpublished data Osmotin Mimics Adiponectin

8. 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 ↑ ↑↓

9. Body Weight Gain DSS+ SAL DSS+ OSM 20 10 0 -10 % body weight gain Water 50 30 10 0 40 20 DSS+ OSM DSS+ SAL Water Percent Hematocrit Osmotin Treatment Reduces Weight Loss And Anemia Associated With Induced Colitis In Mice colitis * * R. Arsenescu

10. Plasma IL-1β (Pro-inflammatory cytokine) DSS+ OSM DSS+ SAL WATER 0 100 200 pg/ml Osmotin Treatment Reduces Inflammation Associated With Induced Colitis In Mice DSS+ OSM DSS+ SAL WATER 0 50 100 150 pg/ml Plasma IL-10 (Anti-inflammatory cytokine) colitis * R. Arsenescu