Journal: PLoS ONE

Article Title: Posttranslational Modification of 6-phosphofructo-1-kinase as an Important Feature of Cancer Metabolism

doi: 10.1371/journal.pone.0019645

Figure Lengend Snippet: Activities of the native PFK1 isolated from rabbit muscle after limited proteolysis by Proteinase K (dark) and untreated native enzyme (light) as measured in a system containing 5 mM citrate. Data are representative of three independent measurements and are presented as means ± standard deviation.

Article Snippet: Human muscle-type PFK1 cDNA (Clone ID2964710) was purchased from Geneservice Ltd ( www.Geneservice.co.uk ).

Techniques: Isolation, Standard Deviation

Journal: PLoS ONE

Article Title: Posttranslational Modification of 6-phosphofructo-1-kinase as an Important Feature of Cancer Metabolism

doi: 10.1371/journal.pone.0019645

Figure Lengend Snippet: Western blots of four tumorigenic cell lines (above) showed the presence of fragments of different lengths, with a fragment of 47 kDa regularly present, while no native 85-kDa PFK1 could be observed. In non-neoplastic cell lines (below) (HEK cells), the native PFK1 forms were predominant, while in normal lymphocytes isolated from peripheral human blood only a single protein band was detected corresponding to 85 kDa native PFK1. In the Western blot of lymphocytes two volumes of cell lysate were applied to the gel: 10 µl (left), and 20 µl (right).

Article Snippet: Human muscle-type PFK1 cDNA (Clone ID2964710) was purchased from Geneservice Ltd ( www.Geneservice.co.uk ).

Techniques: Western Blot, Isolation

Journal: PLoS ONE

Article Title: Posttranslational Modification of 6-phosphofructo-1-kinase as an Important Feature of Cancer Metabolism

doi: 10.1371/journal.pone.0019645

Figure Lengend Snippet: No native PFK1 enzyme was detected in the cells growing in a tissue culture, while in a tumor, a strong signal corresponding to the native enzyme was present. Shorter fragments were detected in both homogenates with a 47 fragment present in individually growing cells and a 45 kDa fragment present in a tumor tissue.

Article Snippet: Human muscle-type PFK1 cDNA (Clone ID2964710) was purchased from Geneservice Ltd ( www.Geneservice.co.uk ).

Techniques:

Journal: PLoS ONE

Article Title: Posttranslational Modification of 6-phosphofructo-1-kinase as an Important Feature of Cancer Metabolism

doi: 10.1371/journal.pone.0019645

Figure Lengend Snippet: In relative specific PFK1 activities detected in the homogenate of the transformant encoding Fragment 9 (□ ) and with native PFK-M enzyme isolated from E.coli transformant (○) are shown, that were measured at increasing concentrations of ATP. In specific PFK1 activities were measured in the homogenate of the transformant encoding Fragment 9 without inhibitor (□), in the presence of 5 mM Na 3 -citrate (◊), and with 5 mM Na-lactate (Δ). All measurements were conducted with 0.5 mM of F6P. Data are representative of at least three independent measurements and are presented as means ± standard deviation.

Article Snippet: Human muscle-type PFK1 cDNA (Clone ID2964710) was purchased from Geneservice Ltd ( www.Geneservice.co.uk ).

Techniques: Isolation, Standard Deviation

Journal: Cancers

Article Title: The J2-Immortalized Murine Macrophage Cell Line Displays Phenotypical and Metabolic Features of Primary BMDMs in Their M1 and M2 Polarization State

doi: 10.3390/cancers13215478

Figure Lengend Snippet: TGFβ stimulation on iμΦ leads to increase of αSMA. ( A ) Gating strategy for the expression of the macrophage marker anti-F4/80-FITC and the myofibroblast marker αSMA-APC by flow cytometry in iμΦ after a 5 day treatment with TGFβ (5 ng/mL). Unstimulated iµФ were used as a control (CTRL). ( B ) Quantification of the αSMA + cell population ( n = 3). Data are means ± SEM. ** p < 0.01.

Article Snippet: Next, the co-expression of macrophage and myofibroblast markers anti-F4/80-FITC and anti-α smooth muscle actin (αSMA)-APC (Clone E184, Abcam) were measured by flow cytometry.

Techniques: Expressing, Marker, Flow Cytometry