Reduction of heart failure by pharmacological inhibition or gene deletion of protein tyrosine phosphatase 1B  Elodie Gomez, Magali Vercauteren, Baptiste Kurtz, Antoine Ouvrard-Pascaud, Paul Mulder, Jean-Paul Henry, Marie Besnier, Aurélie Waget, Rob Hooft Van Huijsduijnen, Michel L. Tremblay, Remy Burcelin, Christian Thuillez, Vincent Richard  Journal of Molecular and Cellular Cardiology  Volume 52, Issue 6, Pages 1257-1264 (June 2012) DOI: 10.1016/j.yjmcc.2012.03.003 Copyright © 2012 Elsevier Ltd Terms and Conditions

Fig. 1 a) Echocardiographic and hemodynamic parameters in normal (n=13), or 2-month CHF mice, either untreated (n=13) or chronically treated with the PTP1B inhibitor AS279 (n=15). * p<0.05 and ** p<0.01 vs. normal mice; † p<0.05 and †† p<0.01 vs. untreated CHF b) Echocardiographic and hemodynamic parameters in normal or 2-month CHF mice, either WT (normal n=11, CHF n=12) or PTP1B−/− (normal n=11, CHF n=9) * p<0.05 and ** p<0.01 vs. WT normal mice; † p<0.05 and †† p<0.01 vs. WT CHF mice. Journal of Molecular and Cellular Cardiology 2012 52, 1257-1264DOI: (10.1016/j.yjmcc.2012.03.003) Copyright © 2012 Elsevier Ltd Terms and Conditions

Fig. 2 a) LV weight, LV weight/tibia length, infarct size and LV collagen density obtained normal (n=13) or CHF mice, either untreated (n=11) or chronically treated with the PTP1B inhibitor AS279 (n=15) (top) b) LV weight, LV weight/tibia length, infarct size and LV collagen density obtained in WT (normal n=17, CHF n=15) or PTP1B−/− mice (normal n=20, CHF n=16). Representative images of collagen density in the four groups are also presented. ** p<0.01 vs. normal mice; † p<0.05 and †† p<0.01 vs. untreated or WT CHF mice. Journal of Molecular and Cellular Cardiology 2012 52, 1257-1264DOI: (10.1016/j.yjmcc.2012.03.003) Copyright © 2012 Elsevier Ltd Terms and Conditions

Fig. 3 a) Effect of CHF and chronic pharmacological PTP1B inhibition on flow-mediated vasodilatation of mesenteric arteries (n=8–9 animals per group). Repeated measures ANOVA revealed statistically significant differences (p<0.01) between normal and CHF and between CHF and CHF+chronic PTP1B inhibition. b) Effect of CHF and chronic pharmacological PTP1B inhibition on the vasodilatory responses of mesenteric arteries to acetylcholine or sodium nitroprusside (SNP) (n=8–9 animals per group). c) Effect of in vitro incubation with the NOS inhibitor L-NNA and the PI3K inhibitor wortmannin of arteries isolated from CHF mice chronically treated with the PTP1B inhibitor AS279 (n=7 for L-NNA and 11 for wortmannin). Repeated measures ANOVA revealed statistically significant differences between base and L-NNA (p<0.01) and between base and wortmannin (p<0.05). d) Representative Western blot data of total eNOS or P Ser1177-eNOS , and mean±SEM P Ser1177-eNOS/eNOS ratio and eNOS (n=6) in mesenteric arteries isolated from normal or CHF mice, either untreated or chronically treated with the PTP1B inhibitor AS279, and exposed to flow at 200μl/min for 2min. and 30sec. Journal of Molecular and Cellular Cardiology 2012 52, 1257-1264DOI: (10.1016/j.yjmcc.2012.03.003) Copyright © 2012 Elsevier Ltd Terms and Conditions

Fig. 4 a) Flow-mediated vasodilatation of mesenteric arteries obtained from WT (n=16) or PTP1B−/− (n=8) normal mice (in the absence of CHF). Repeated measures ANOVA revealed statistically significant differences (p<0.01) between WT and PTP1B−/−. b) Flow-mediated vasodilatation of mesenteric arteries obtained from WT (n=10) or PTP1B−/− mice (n=12) with CHF. Repeated measures ANOVA revealed statistically significant differences (p<0.01) between WT and PTP1B−/− CHF mice. c) Effect of acute in vitro incubation with the PTP1B inhibitor AS179 on flow-mediated vasodilatation of mesenteric arteries isolated from PTP1B−/− CHF mice (n=7 ). d) Representative PCR experiments showing PTP1B expression in the liver, lung, RV, LV, and small mesenteric artery (MA) from WT or PTP1B−/− normal mice. Journal of Molecular and Cellular Cardiology 2012 52, 1257-1264DOI: (10.1016/j.yjmcc.2012.03.003) Copyright © 2012 Elsevier Ltd Terms and Conditions

Fig. 5 mRNA expression of NOS isoforms (normalized to 18S) in mesenteric arteries (left) or hearts (right) from WT or PTP1B−/− mice, either normal or CHF (n=6 per group). Journal of Molecular and Cellular Cardiology 2012 52, 1257-1264DOI: (10.1016/j.yjmcc.2012.03.003) Copyright © 2012 Elsevier Ltd Terms and Conditions

Fig. 6 a) Glucose turnover rate (ml/kg·min) assessed in euglycemic hyperinsulinemic clamp studies performed in normal mice (n=16), or CHF mice, either WT (n=9) or PTP1B−/− (n=17). * p<0.05 vs. CHF WT. b) Vasodilatory responses to insulin of mesenteric arteries obtained from normal (n=6) or CHF mice, either WT (n=7) or PTP1B−/− (n=5). Repeated measures ANOVA revealed statistically significant differences between normal and CHF mice and between WT CHF and PTP1B−/− CHF mice. Journal of Molecular and Cellular Cardiology 2012 52, 1257-1264DOI: (10.1016/j.yjmcc.2012.03.003) Copyright © 2012 Elsevier Ltd Terms and Conditions