goat anti ampkα1 polyclonal antibody  (R&D Systems)

Journal: The Journal of Biological Chemistry

Article Title: 5′-AMP-activated Protein Kinase (AMPK) Supports the Growth of Aggressive Experimental Human Breast Cancer Tumors *

doi: 10.1074/jbc.M114.576371

Figure Lengend Snippet: A and B, transcript levels of AMPKα1 (PRKAA1) and AMPKα2 (PRKAA2) in shRNA control and AMPKα1/2 KD MDA-MB-231 cells. Numerical values refer to % (KD Signal/Control Signal) ± % combined S.D. (n = 2 replicates). C, AMPKα1 and AMPKα2 protein levels in shRNA control and AMPKα1/2 KD MDA-MB-231 cells. Numerical values refer to % (normalized KD signal/normalized control signal) ± % combined S.D. (n = 2 replicates). AMPKα signals were normalized to the corresponding MARK2 signals (signals are intensity values (thousand pixels) obtained by image analysis). D, total AMPKα (AMPKα1/2), ACC1, and phospho-Ser-79 ACC1 (P-ACC1) protein levels in shRNA control and AMPKα1/2 KD MDA-MB-231 cells. Numerical values refer to % (normalized KD signal/normalized control signal) ± % combined S.D. (n = 2, replicates). AMPKα signals were normalized to the corresponding MARK2 signals (signals are intensity values (thousand pixels) obtained by image analysis).

Article Snippet: Materials Goat polyclonal anti-AMPKα1 and -α2 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA; catalog nos. sc-19128 and sc-19131, respectively).

Techniques: shRNA, Control

Journal: The Journal of Biological Chemistry

Article Title: 5′-AMP-activated Protein Kinase (AMPK) Supports the Growth of Aggressive Experimental Human Breast Cancer Tumors *

doi: 10.1074/jbc.M114.576371

Figure Lengend Snippet: A, effect of an AMPKα1/2 KD on the growth of orthotopic MDA-MB-231 xenografts in nude mice (10 implanted mice/group). Mean tumor volume data were analyzed by one-way analysis of variance (p < 0.0001). The asterisks refer to p values for statistically significant differences (p < 0.05; Tukey-Kramer test) at the indicated days of tumor growth: *, day 40, p < 0.001; **, day 36, p < 0.05 (shRNA control group, top curve/diamonds; AMPKα1/2 KD group, bottom curve/squares). Error bars, ±S.D. B, in vitro proliferation/survival curves for shRNA control and AMPKα1/2 KD MDA-MB-231 cells obtained by using an Alamar Blue proliferation/viability assay. Raw fluorescence data were fitted by nonlinear regression to a sigmoidal growth equation. Error bars, ±S.D. (n = 3 replicates). C, survival curves from a lung colonization assay experiment involving control and AMPKα1/2 KD MDA-MB-231 cells. MDA-MB-231 cells were injected into the tail vein of beige-nude mice (2 × 106 cells/injection; 15 mice/group). Survival was determined by the Kaplan-Meier method (p < 0.0001 for median survival). Top curve (triangles), AMPKα1/2 KD group; bottom curve (squares), shRNA control group.

Article Snippet: Materials Goat polyclonal anti-AMPKα1 and -α2 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA; catalog nos. sc-19128 and sc-19131, respectively).

Techniques: shRNA, Control, In Vitro, Viability Assay, Fluorescence, Injection

Journal: The Journal of Biological Chemistry

Article Title: 5′-AMP-activated Protein Kinase (AMPK) Supports the Growth of Aggressive Experimental Human Breast Cancer Tumors *

doi: 10.1074/jbc.M114.576371

Figure Lengend Snippet: A, in vitro growth curves for shRNA control and AMPKα1/2 KD MDA-MB-231 cells cultured in high and low glucose-containing medium (25 or 1 mm glucose in DMEM + 10% FBS; data obtained by cell counting). Error bars, ±S.D. (n = 3 replicates). Top curves (solid lines), 25 mm glucose; bottom curves (broken lines), 1 mm glucose. B, effect of AMPKα1/2 KD on the uptake of glucose (top) and glutamine (bottom) by MDA-MB-231 cells cultured under normoxia (5% CO2 + air). Cells were given 2-deoxy-d-[2,6-3H]glucose or l-[3,4-3H]glutamine for 5 min (see under “Experimental Procedures” for details). Glucose uptake: shRNA control cells, 129 ± 22 pmol/mg total protein/5 min; AMPKα1/2 KD cells, 106 ± 11 pmol/mg total protein/5 min; ±S.D., n = 3. Glutamine uptake: shRNA control cells, 10,411 ± 1,034 cpm/mg total protein/5 min; AMPKα1/2 KD cells, 9320 ± 314 pmol/mg total protein/5 min; ±S.D., n = 3.

Article Snippet: Materials Goat polyclonal anti-AMPKα1 and -α2 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA; catalog nos. sc-19128 and sc-19131, respectively).

Techniques: In Vitro, shRNA, Control, Cell Culture, Cell Counting

Journal: The Journal of Biological Chemistry

Article Title: 5′-AMP-activated Protein Kinase (AMPK) Supports the Growth of Aggressive Experimental Human Breast Cancer Tumors *

doi: 10.1074/jbc.M114.576371

Figure Lengend Snippet: Metabolic profiling of shRNA control and AMPKα1/2 KD MDA-MB-231 cells cultured with a d-[1,2-13C2]glucose tracer in high and low glucose-containing medium (25 or 1 mm glucose in DMEM +10% FBS). Histograms show the effect of AMPKα1/2 KD on glucose metabolism through the following pathways. A, 13C-labeling patterns of selected molecules of core glucose metabolism evaluated using [13C2]glucose as a tracer. B, oxidation to CO2 in the PPC and the TCA cycle; C, production of lactate by aerobic glycolysis and the PPC; D–F, production of RNA ribose by triose recycling (D), the PPC (E), the oxidative PPC (F), and the nonoxidative PPC (G); production of palmitate from all pathways (H) and from acetyl-CoA from the TCA cycle (I). Error bars, ±S.D. (n = 3 plates/cell line).

Article Snippet: Materials Goat polyclonal anti-AMPKα1 and -α2 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA; catalog nos. sc-19128 and sc-19131, respectively).

Techniques: shRNA, Control, Cell Culture, Labeling

Journal: The Journal of Biological Chemistry

Article Title: 5′-AMP-activated Protein Kinase (AMPK) Supports the Growth of Aggressive Experimental Human Breast Cancer Tumors *

doi: 10.1074/jbc.M114.576371

Figure Lengend Snippet: Effect of AMPKα1/2 KD on the oxygen consumption rate (OCR) (A) and proton production rate (PPR) (B) of MDA-MB-231 cells cultured under normoxia (5% CO2 + air). Open arrow, addition of the ATP synthase inhibitor oligomycin to inhibit oxidative phosphorylation. Closed arrow, addition of the uncoupling agent/protonophore (carbonyl cyanide m-chlorophenyl hydrazone) to uncouple the respiratory chain from ATP synthesis. Error bars, ±S.D.

Article Snippet: Materials Goat polyclonal anti-AMPKα1 and -α2 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA; catalog nos. sc-19128 and sc-19131, respectively).

Techniques: Cell Culture, Phospho-proteomics

Journal: The Journal of Biological Chemistry

Article Title: 5′-AMP-activated Protein Kinase (AMPK) Supports the Growth of Aggressive Experimental Human Breast Cancer Tumors *

doi: 10.1074/jbc.M114.576371

Figure Lengend Snippet: Metabolic profiling of shRNA control and AMPKα1/2 KD MDA-MB-231 orthotopic tumors with a d-[13C6]glucose tracer. A, 13C-labeling patterns of selected molecules of core glucose metabolism evaluated using [13C6]glucose as a tracer. B, growth curves for the orthotopic MDA-MB-231 tumors used for the study (30 implanted mice/group). Mean tumor volume data were analyzed by one-way analysis of variance (p < 0.0001). The asterisks refer to p values (p < 0.05; Tukey-Kramer test) for statistically significant differences in mean tumor volumes (p < 0.050) at the indicated days of tumor growth: *, day 21, p < 0.001; **, day 18, p < 0.001 (shRNA control group, top curve/diamonds; AMPKα1/2 KD group, bottom curve/squares). Error bars, ±S.D. Histograms show the effect of AMPKα1/2 KD on core glucose metabolism in tumor tissue (during ≤90 min; left-to-right, 30-, 60-, and 90-min observation times; five tumors/time) through the following pathways: C, oxidation to CO2 in the PPC and the TCA cycle; D and E, production of lactate by glycolysis and the PPC; F, production of lactate by the oxidative PPC relative to glycolysis; production of RNA ribose by all pathways (G) and the nonoxidative PPC (H); production of palmitate from all pathways (I and J) and from acetyl-CoA from the TCA cycle (K). Tumor tissue and plasma samples were obtained on day 21 at 30-, 60-, and 90-min after i.p. injection of the tracer (15 mice from each group were used for metabolic profiling; 5 mice/time point; see “Experimental Procedures” for details). Error bars, ±S.D.

Article Snippet: Materials Goat polyclonal anti-AMPKα1 and -α2 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA; catalog nos. sc-19128 and sc-19131, respectively).

Techniques: shRNA, Control, Labeling, Clinical Proteomics, Injection

Journal: The Journal of Biological Chemistry

Article Title: 5′-AMP-activated Protein Kinase (AMPK) Supports the Growth of Aggressive Experimental Human Breast Cancer Tumors *

doi: 10.1074/jbc.M114.576371

Figure Lengend Snippet: Metabolic profiling of pancreas and liver from mice used for the study detailed in Fig. 6. Histograms show the effect of AMPKα1/2 KD on glucose metabolism in pancreas and liver tissue through the following pathways: A, production of RNA ribose by all pathways; and B, production of lactate by the oxidative PPC relative to glycolysis. C, enrichment of 13C from the d-[13C6]glucose tracer in plasma glucose. Error bars, ±S.D.

Article Snippet: Materials Goat polyclonal anti-AMPKα1 and -α2 antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA; catalog nos. sc-19128 and sc-19131, respectively).

Techniques: Clinical Proteomics