Journal: Nature medicine

Article Title: Activation of AMPKα2 in adipocytes is essential for nicotine-induced insulin resistance in vivo

doi: 10.1038/nm.3826

Figure Lengend Snippet: Adipose AMPKα2 is required for the nicotine-dependent inhibitory effects on weight gain and insulin signaling in mice. Nic or Veh was perfused in control (Con, black), Prkaa1 Δad (brown) and Prkaa2 Δad (purple) mice. ( a ) Phosphorylated AMPK at Thr172 (pAMPK) and phosphorylated ACC (pACC) expression in WAT; n = 8 each. ( b ) Genotypes of adipose-specific knockout mice and the expressions of AMPKα, AMPKα1, and AMPKα2 in isolated adipocytes; n = 8 each. ( c ) Body weight in mice perfused with Veh or Nic; n = 8–9 each. ( d ) Representative H&E-stained WAT sections ( n = 10 section each) and the diameters of adipocytes in WAT (Scale bar, 50 µm); n = 8 each. ( e ) Serum FFA levels after overnight fasting (Fasted) or insulin perfusion at 40 min during ITT; n = 8 each. ( f ) IPGTT in mice treated with Veh or Nic; n = 8–9 each. ( g ) Irs1, pIrs1-Ser307 (pIrs1), pAkt-Ser473 (pAkt), p38, and JNK signaling in WAT of mice injected with saline or insulin (1 unit kg −1 ); n = 8 each. Significance determined by Student’s t -test ( a,d ), one-way ANOVA with repeated measures for the inter-assay evaluations ( c,f ), one-way ANOVA with Bonferroni’s post-hoc test ( e,g ) and * P < 0.05. All values are means ± SEM.

Article Snippet: Antibodies against AMPK-α (2532), AMPK-α1 (2795), AMPK-α2(2757), pAMPK (Thr172, 2535), pACC (Ser79, 3661), ACC (3662), p38 (9212), pp38 (4631), JNK (9258), pJNK (9251), pIRS1 (Ser307, 2381), pIRS1 (Ser636/639, 2388), pIRS1 (Ser789, 2389), p-mTOR (Ser2448, 5536), mTOR (2983), pS6K (Thr389, 9324), S6K (2708), pRaptor (Ser789, 2083), Raptor (2280), pThr (9386) and histone H3 (9717) were purchased from Cell Signaling Technology.

Techniques: Control, Expressing, Knock-Out, Isolation, Staining, Injection, Saline, Inter Assay

Journal: Nature medicine

Article Title: Activation of AMPKα2 in adipocytes is essential for nicotine-induced insulin resistance in vivo

doi: 10.1038/nm.3826

Figure Lengend Snippet: MKP1 reduction is required for nicotine-mediated IR and lipolysis. ( a–g ) WT and Mkp1 −/− mice were treated with Veh or Nic. ( a ) Body weight changes and fat mass in mice treated with Veh or Nic; n = 7–8 each. ( b ) In vivo lipolysis. Plasma glycerol concentration measured at 0, 15, 60, 120 min; n = 6 each. ( c,d ) IPGTT ( c ) and ITT ( d ) data in mice; n = 7–8 each. ( e,f ) The glucose infusion rate (GIR) ( e ) and the hepatic glucose production (HGP), the rate of the disappearance (R d ) and in tibialis anterior and soleus muscles or eWAT the glucose metabolic index (R g ) ( f ) during a hyperinsulinemic-euglycemic clamp ( n = 6 each). ( g ) MKP1, pIrs1- Ser307, Irs1 and p38 signaling in WAT of mice of the indicated genotype; n = 6 each. ( h ) AchRα7 expression in isolated adipocytes, hepatocytes, β-cells, and myocytes after treatment with Nic or Veh; n = 6 each. ( i ) Detection of pAMPK, AMPK, MKP1, and Irs1 expression in WAT in smokers and nonsmokers; n = 12 each. ( j ) Oral glucose tolerance test (OGTT) (left), the insulin levels during OGTT (right) and the areas under the curve (AUC); n = 12 each. Significance determined by one-way ANOVA with Bonferroni’s post-hoc test ( a,f,g,j ), one-way ANOVA with repeated measures for the inter-assay evaluations ( b,c,d,e ), Student’s t -test ( h,i ), * P < 0.05 Veh vs.Nic, and # P < 0.05 WT vs. Mkp1 −/− mice. All values are means ± SEM.

Article Snippet: Antibodies against AMPK-α (2532), AMPK-α1 (2795), AMPK-α2(2757), pAMPK (Thr172, 2535), pACC (Ser79, 3661), ACC (3662), p38 (9212), pp38 (4631), JNK (9258), pJNK (9251), pIRS1 (Ser307, 2381), pIRS1 (Ser636/639, 2388), pIRS1 (Ser789, 2389), p-mTOR (Ser2448, 5536), mTOR (2983), pS6K (Thr389, 9324), S6K (2708), pRaptor (Ser789, 2083), Raptor (2280), pThr (9386) and histone H3 (9717) were purchased from Cell Signaling Technology.

Techniques: In Vivo, Clinical Proteomics, Concentration Assay, Muscles, Expressing, Isolation, Inter Assay