phospho ser79 acc pacc  (Cell Signaling Technology Inc)

Journal: Biofactors (Oxford, England)

Article Title: Diverse Inhibitors of De Novo Purine Synthesis Promote AICAR ‐Induced AMPK Activation and Glucose Uptake in L6 Myotubes

doi: 10.1002/biof.70037

Figure Lengend Snippet: Mycophenolate mofetil, alanosine, trimetrexate, and sulfamethoxazole promote AICAR‐induced AMPK activation and glucose uptake in L6 myotubes. L6 myotubes were incubated in nucleoside‐ and serum‐free MEMα for 24 h and treated with 5 μM mycophenolate mofetil (MMF), 5 μM alanosine (ALA), MMF + ALA, 50 μM mercaptopurine (MP), 20 μM trimethoprim (TMP), 500 μM sulfamethoxazole (SMX), 5 μM trimetrexate (TMX), 5 μM methotrexate (MTX) or vehicle (Control) for the last 16 h of these 24 h and with 1 mM AICAR or vehicle for the last 60 min of these 24 h for analysis of protein phosphorylation or with 2 mM AICAR or vehicle for the last 5 h of these 24 h for analysis of glucose uptake. Following the treatment, cells were analyzed for phospho AMPKα Thr172 (pAMPK) and AMPKα (AMPK) (A, B) and phospho ACC Ser79 (pACC) and ACC (C, D) with immunoblotting or for glucose uptake with 2‐deoxy‐glucose (2DG) uptake assay (E, F). 2DG uptake was expressed in pmol of 2DG/min/mg of total proteins. Graphs show means with SD ( n = 4–6). Images show representative blots. Numbers next to blots indicate molecular weight (in kDa) of the first marker below and/or above the bands on the blots. * p < 0.05 versus respective (vehicle or AICAR) control, # p < 0.05 AICAR control versus vehicle control; one‐way ANOVA with Bonferroni's test. AU, arbitrary units.

Article Snippet: pACC Ser79 , CST #3661 , Rabbit , pAb , 1:1000.

Techniques: Activation Assay, Incubation, Control, Phospho-proteomics, Western Blot, Molecular Weight, Marker

Journal: Biofactors (Oxford, England)

Article Title: Diverse Inhibitors of De Novo Purine Synthesis Promote AICAR ‐Induced AMPK Activation and Glucose Uptake in L6 Myotubes

doi: 10.1002/biof.70037

Figure Lengend Snippet: Effect of allopurinol on AMPK, insulin signaling, and glucose uptake in L6 myotubes. L6 myotubes were incubated in nucleoside‐ and serum‐free MEMα for 24 h and treated with 100 μM allopurinol (ALO), 5 μM methotrexate (MTX) or vehicle (Control) for the last 16 h of these 24 h and with 1 mM AICAR or vehicle for the last 60 min of these 24 h or with 120 nM insulin or vehicle for the last 20 min of these 24 h for analysis of protein phosphorylation (A‐F) or with 2 mM AICAR or vehicle for the last 5 h of these 24 h or with 120 nM insulin or vehicle for the last 60 min of these 24 h for analysis of glucose uptake (G). Following the treatment, cells were analyzed for phospho AMPKα Thr172 (pAMPK) and AMPKα (AMPK) (A), phospho ACC Ser79 (pACC) and ACC (B), phospho Akt Ser473 (pAkt) and Akt (C, D) and phospho ERK1/2 Thr202/Tyr204 (pERK1/2) and ERK1/2 (E, F) with immunoblotting or for glucose uptake (G) with a 2‐deoxy‐glucose (2DG) uptake assay. For the analysis of phospho Akt and phospho ERK1/2, we recorded one short‐exposure and one long‐exposure image. The short‐exposure image was used for the analysis of insulin‐induced phosphorylation (C, E), while the long‐exposure image was used for the analysis of basal phosphorylation (D, F). 2DG uptake was expressed in pmol of 2DG/min/mg of total proteins. Graphs show means with SD ( n = 4). Images show representative blots. Numbers next to blots indicate molecular weight (in kDa) of the first marker below and/or above the bands on the blots. * p < 0.05 versus respective (vehicle, AICAR or insulin) control, # p < 0.05 AICAR or insulin control versus vehicle control; one‐way ANOVA with Bonferroni's test. AU, arbitrary units.

Article Snippet: pACC Ser79 , CST #3661 , Rabbit , pAb , 1:1000.

Techniques: Incubation, Control, Phospho-proteomics, Western Blot, Molecular Weight, Marker