Journal: Journal of applied physiology (Bethesda, Md. : 1985)

Article Title: Exercise effects on γ3-AMPK activity, Akt substrate of 160 kDa phosphorylation, and glucose uptake in muscle of normal and insulin-resistant female rats.

doi: 10.1152/japplphysiol.00533.2021

Figure Lengend Snippet: Figure 2. A: phosphorylated (p) AMP-activated protein kinase (AMPK)aThr172/AMPKa in epitrochlearis muscles immediately postexercise (IPEX). IPEX vs. sedentary (SED) within each diet (P < 0.001). HFD, high-fat diet; LFD, low-fat diet. B: phosphorylated Akt substrate of 160 kDa (AS160)Ser704/AS160 in epi- trochlearis muscles at IPEX. IPEX vs. SED within each diet (P < 0.001 for LFD group; P < 0.01 for HFD group). C: phosphorylated acetyl CoA carboxylase (ACC)Ser79/ACC in epitrochlearis muscles at IPEX. IPEX vs. SED within each diet (P < 0.01). Data were analyzed by 2-way analysis of variance. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 12 rats per treatment group. The representative blots are aligned with the group identification in the bar graph.

Article Snippet: Anti-phospho Akt Ser473 (pAktSer473; #9271), anti-phospho Akt Thr308 (pAktThr308; #13038), anti-Akt (#4691), anti-phospho AS160 Thr642 (pAS160Thr642; #8881), anti-phospho AS160 Ser588 (pAS160Ser588; #8730), anti-phospho AMPKa Thr172 (pAMPKaThr172; #2531), anti-AMPK-a (AMPKa; #5831), antiacetyl CoA carboxylase (ACC; #3676), anti-phospho ACC Ser79 (pACCSer79; #3661), anti-hexokinase II (HKII; #2867), and anti-rabbit IgG horseradish peroxidase (HRP) conjugate (#7074) were from Cell Signaling Technology (Danvers, MA).

Techniques: Muscles

Journal: Journal of applied physiology (Bethesda, Md. : 1985)

Article Title: Exercise effects on γ3-AMPK activity, Akt substrate of 160 kDa phosphorylation, and glucose uptake in muscle of normal and insulin-resistant female rats.

doi: 10.1152/japplphysiol.00533.2021

Figure Lengend Snippet: Figure 6. A: phosphorylated (p) Akt substrate of 160 kDa (AS160)Ser704/AS160 in paired epitrochlearis muscles incubated ± 100 mU/mL insulin at 3 h post- exercise (3hPEX). HFD, high-fat diet; LFD, low-fat diet; SED, sedentary. 3hPEX vs. SED within each diet with no insulin (P < 0.01 for LFD group; P < 0.05 for HFD group); †3hPEX vs. SED within each diet with insulin (P < 0.05 for LFD group; P < 0.01 for HFD group). B: delta-insulin pAS160Ser704/AS160. C: pAS160Ser588/AS160 in paired epitrochlearis muscles incubated ± 100 mU/mL insulin at 3hPEX. 3hPEX vs. SED within each diet with no insulin (P < 0.01 for LFD group; P < 0.05 for HFD group); †3hPEX vs. SED within LFD group with insulin (P < 0.05). D: delta-insulin pAS160Ser588/AS160. E: pAS160Thr642/ AS160 in paired epitrochlearis muscles incubated ± 100 mU/mL insulin at 3hPEX. †3hPEX vs. SED within each diet with insulin (P < 0.05). F: delta-insulin pAS160Thr642/AS160. Data were analyzed by 2-way analysis of variance within each insulin level (without or with insulin) or for delta-insulin values. Delta- insulin values= value with insulin value without insulin from paired muscles. Tukey post hoc analysis was performed to identify significant differences. Values are means ± SD; n = 10 rats per treatment group. The representative blots are aligned with the group identification in the bar graph.

Article Snippet: Anti-phospho Akt Ser473 (pAktSer473; #9271), anti-phospho Akt Thr308 (pAktThr308; #13038), anti-Akt (#4691), anti-phospho AS160 Thr642 (pAS160Thr642; #8881), anti-phospho AS160 Ser588 (pAS160Ser588; #8730), anti-phospho AMPKa Thr172 (pAMPKaThr172; #2531), anti-AMPK-a (AMPKa; #5831), antiacetyl CoA carboxylase (ACC; #3676), anti-phospho ACC Ser79 (pACCSer79; #3661), anti-hexokinase II (HKII; #2867), and anti-rabbit IgG horseradish peroxidase (HRP) conjugate (#7074) were from Cell Signaling Technology (Danvers, MA).

Techniques: Muscles, Incubation

Journal: Frontiers in Physiology

Article Title: Impact of PCSK9 on CTRP9-Induced Metabolic Effects in Adult Rat Cardiomyocytes

doi: 10.3389/fphys.2021.593862

Figure Lengend Snippet: Impact of PCSK9 on CTPR9-induced effects on glucose metabolism. Adult rat cardiomyocytes were incubated in the presence of LacZ-containing supernatant (control), murine wild-type PCSK9 or GOF PCSK9 (each 0.5 μg/ml) in serum-free medium for 24 h and subsequently treated with CTRP9 (4 μg/ml) for the indicated times. (A) Phosphorylation of Akt and AS160 was analyzed after 20 min CTRP9 stimulation by Western Blotting. Representative Western blots and densitometry of protein data are shown. Total Akt and total AS160 as well as GAPDH served as a loading control. (B) Translocation of GLUT-4 from the cytosol to the plasma membrane was analyzed after 30 min CTRP9 stimulation. Representative Western blots and densitometry of protein data are shown. Na-K-ATPase served as a marker for membrane fraction and GAPDH as a marker for the cytosol. (C) Glucose uptake was measured after 10 min stimulation with CTRP9. (D) Changes in GLUT-4, hexokinase and PFK1 mRNA expression after 24 h stimulation with CTRP9 were analyzed by Real-time PCR. All data are mean ± SEM, 4 independent experiments, n = 8 per group, * p < 0.05.

Article Snippet: Following blocking, filters were incubated with antibodies directed against, phospho-AMPK (Thr172), alpha-AMPK, phospho-Akt (Thr308), total-Akt, phospho-AS160 (Thr642), total-AS160 (all Cell Signaling Technology), PCSK9 (Abcam), ND1 (Abcam), Cox I (Thermo Fisher Scientific), Na-K-ATPase and GAPDH (both Abcam).

Techniques: Incubation, Control, Phospho-proteomics, Western Blot, Translocation Assay, Clinical Proteomics, Membrane, Marker, Expressing, Real-time Polymerase Chain Reaction