uthor manuscript; available in PMC 2012 January 24. NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript Figure 6. Modulation of p110γ Lipid Kinase Activity by LY335979 167465-36-3 PKA Affects β-AR Density Myocardial β-AR density in wild-type , p110β kinase-dead , p110γ knockout , and p110γ kinase-dead mice. Myocardial β-AR density of p110γ+/+ and p110γKD/KD mice subjected to sham operation or to aortic constriction for 20 weeks. Myocardial PtdIns P3 levels in hearts obtained from sham-operated mice or from 20 week TAC-treated wild-type or p110γKD/KD mice. Lipid kinase activity of p110γ immunoprecipitated from myocardial tissue lysates of sham-operated or 20 week TAC-treated mice. Values were obtained by quantitative densitometry and normalized over control.
Coimmunoprecipitation of p110γ and PKA C from myocardial lysates from mice subjected to pressure overload for 20 weeks or to sham operation. After quantitative densitometry, p110γ-bound PKA was expressed as the ratio of coimmunoprecipitated Alvespimycin HSP-90 inhibitor PKA C over immunoprecipitated p110γ. Total levels of the indicated proteins in myocardial tissue lysates from sham-operated mice or from mice subjected to pressure overload for 20 weeks. mRNA levels of p110γ, p101, and p84/p87 in hearts from sham-operated mice or mice subjected to 20 weeks of TAC. Coimmunoprecipitation of p110γ with p84/87 from myocardial lysates of mice subjected to pressure overload for 20 weeks or to sham operation. Coimmunoprecipitation of p110γ with p101 from myocardial lysates of mice subjected to pressure overload for 20 weeks or to sham operation.
In β-AR density measurements , receptor density is expressed as the Bmax after saturation binding using 125I-labeled cyanopindolol ligand. A representative assay is presented in �? For all bar graphs, values represent mean _ SEM of a minimum of four independent experiments or six mice per group. *p 0.05, **p 0.01, ***p 0.001. See also Figure S7. Perino et al. Page 17 Mol Cell. Author manuscript; available in PMC 2012 January 24. NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript Figure 7. p110γ Inhibition Restores β-AR Density and Cardiac Function in Heart Failure Immunohistochemistry for p110γ in human hearts from healthy control patients or patients with aortic stenosis. Bar graph is 25 μm. Representative images are presented.
Myocardial β-AR density of wild-type mice with heart failure treated with selective p110γ inhibitor AS605240 or vehicle. Left ventricular fractional shortening of wild-type mice with heart failure treated with AS605240 or vehicle or of p110γKD/KD mice subjected to aortic constriction for 20 weeks. Representative M-mode echocardiographic snapshots are presented. For all bar graphs, values represent mean _ SEM of eight mice per group. *p 0.05, **p 0.01. Schematic representation of the diverse function of p110γ and p110β in cardiac physiology and pathology. See also Figure S7. Perino et al. Page 18 Mol Cell. Author manuscript; available in PMC 2012 January 24. NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript Blockade of class IB phosphoinositide-3 kinase ameliorates obesity-induced inflammation and insulin resistance Naoki Kobayashia, Kohjiro Uekia,b,1, Yukiko Okazakia, Aya Iwanea, Naoto Kubotaa,b,c, Mitsuru Ohsugia, Motoharu Awazawaa,c, Masatoshi Kobayashia, Takayoshi Sasakoa, Kazuma Kanekoa, Miho Suzukia, Yoshitaka Nishikawaa, Kazuo Haraa, Kotaro Yoshimurad, Isao Koshimad, Susumu Goyamae, Koji Murakamif, Junko Sasakig, Ryozo Nagaih, Mineo Kurokawae, Takehiko Sasakig, and Takashi Kadowakia,b,c,1 aDepartment of Metabolic Diseases, dDepartment of Plastic Sur