rabbit anti mouse p akt2 s474 d3h2  (Cell Signaling Technology Inc)

Journal: EMBO Reports

Article Title: Liver ChREBP deficiency inhibits fructose-induced insulin resistance in pregnant mice and female offspring

doi: 10.1038/s44319-024-00121-w

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Mouse anti-AKT , Cell Signaling Technology , Cat# 5239S.

Techniques: Isolation, Recombinant, Sequencing, Software, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: Stable knockdown or overexpression of AKT isoforms in HCC cell lines. Knockdown of AKT isoforms was conducted by lentiviral transduction with AKT-isoform-specific shRNAs in Hep3B ( A ) or Huh7 ( B ) cells. Overexpression of AKT1, AKT2 and AKT3 isoforms upon transduction with constitutively activated AKT lentiviral vectors in Hep3B ( C ) or Huh7 ( D ) cells. Protein expression levels of AKT isoforms were examined by Western blot analysis using AKT-isoform-specific antibodies. The experiments were repeated three times and the representative results are shown.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Knockdown, Over Expression, Transduction, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: Effect of AKT1, AKT2 and AKT3 silencing on oxygen metabolism in the Hep3B cell line. ( A ) Knockdown of AKT1, AKT2 and AKT3 reduced the oxygen consumption rate (OCR) in HCC cell lines Hep3B. ( B ) Summary statistics of Basal rate of respiration. ( C ) Summary statistics of spare respiratory capacity. ( D ) Summary statistics of oligomycin-sensitive respiration (ATP production). ( E ) Summary statistics of uncoupled respiration (maximal respiratory capacity). The experiments were repeated six times and the pooled results are shown. p -values were calculated with ordinary one-way ANOVA with Dunnett’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Knockdown, Comparison

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: Effect of AKT1 and AKT2 silencing on the oxygen metabolism in Huh7 cell lines. ( A ) Knockdown of AKT1 and AKT2 reduced the oxygen consumption rate (OCR) in the HCC cell line Huh7. ( B ) Summary statistics of Basal rate of respiration. ( C ) Summary statistics of spare respiratory capacity. ( D ) Summary statistics of oligomycin sensitive respiration (ATP production). ( E ) Summary statistics of uncoupled respiration (maximal respiratory capacity). The experiments were repeated six times and the pooled results are shown. p -values were calculated with ordinary one-way ANOVA with Dunnett’s multiple comparison test. ** p < 0.01, *** p < 0.001.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Knockdown, Comparison

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: Effect of AKT1, AKT2 and AKT3 activation on the oxygen metabolism of the Hep3B cell lines. ( A ) Overexpression of constantly activated AKT1, AKT2 and AKT3 increased the oxygen consumption rate (OCR) in HCC cell lines Hep3B. ( B ) Summary statistics of Basal rate of respiration in Hep3B. ( C ) Summary statistics of spare respiratory capacity in Hep3B. ( D ) Summary statistics of oligomycin-sensitive respiration (ATP production) in Hep3B. ( E ) Summary statistics of uncoupled respiration (maximal respiratory capacity) in Hep3B. The experiments were repeated six times in Hep3B cells and the pooled results are shown. p -values were calculated with ordinary one-way ANOVA with Dunnett’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Activation Assay, Over Expression, Comparison

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: Effect of AKT1, AKT2 and AKT3 activation on the oxygen metabolism of the Huh7 cell lines. ( A ) Overexpression of constantly activated AKT1, AKT2 and AKT3 increased the oxygen consumption rate (OCR) in HCC cell lines Huh7. ( B ) Summary statistics of Basal rate of respiration in Huh7. ( C ) Summary statistics of spare respiratory capacity in Huh7. ( D ) Summary statistics of oligomycin sensitive respiration (ATP production) in Huh7. ( E ) Summary statistics of uncoupled respiration (maximal respiratory capacity) in Huh7. The experiments were repeated five times in Huh7 cells and the pooled results are shown. p -values were calculated with ordinary one-way ANOVA with Dunnett’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Activation Assay, Over Expression, Comparison

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: Knockdown of AKT1, AKT2 and AKT3 suppresses ECAR in HCC cell lines. ( A , D ) Extracellular acidification rate (ECAR) measured by extracellular flux analysis. ( B , E ) Summary statistics of basal ECAR. ( C , F ) Summary statistics of stress ECAR. The experiments were repeated six times and the pooled results are shown. p -values were calculated with ordinary one-way ANOVA with Dunnett’s multiple comparison test *** p < 0.001.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Knockdown, Comparison

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: The effects of AKT1, AKT2 and AKT3 on lactic acid production in HCC cells were evaluated using the lactate assay. ( A ) Stable knockdowns of AKT1, AKT2 and AKT3 were generated in ( A ) Hep3B and ( B ) Huh7 cells. Thereafter, the cells were cultured in lactate-free medium for the indicated times and lactate levels were determined. The experiments were repeated three times and the pooled results are shown. p -values were calculated with ordinary one-way ANOVA with Dunnett’s multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: Lactate Assay, Generated, Cell Culture, Comparison

Journal: International Journal of Molecular Sciences

Article Title: All Three AKT Isoforms Can Upregulate Oxygen Metabolism and Lactate Production in Human Hepatocellular Carcinoma Cell Lines

doi: 10.3390/ijms25042168

Figure Lengend Snippet: Model of the regulation of oxygen consumption and lactate production by the three AKT isoforms. AKT1, AKT2 and AKT3 increase oxygen consumption in HCC cells by a molecular mechanism leading to dephosphorylation of PDH at the inhibitory regulatory Ser232. The upregulation (blue arrows) of metabolic changes compared with control cells are shown.

Article Snippet: Thereafter, the proteins were blotted onto a nitrocellulose membrane and incubated with specific primary antibodies against AKT1 (#2938S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT2 (#5239S, Cell Signaling Technology Inc., Danvers, MA, USA), AKT3 (#8018S, Cell Signaling Technology Inc., Danvers, MA, USA), PDH (#2784S, Cell Signaling Technology Inc., Danvers, MA, USA), p-PDH Ser232 (#15289S, Cell Signaling Technology Inc., Danvers, MA, USA) and GAPDH (#G0622, Santa Cruz Biotechnology Inc., Dallas, TX, USA).

Techniques: De-Phosphorylation Assay, Control