Labeling:Article Title: TET2 loss promotes premalignant survival and clonal selection in MYC-driven B cell lymphoma
Article Snippet: .. Labeling was stopped by adding 4 volumes of cold complete B cell medium (DMEM medium (Sigma-Aldrich, D6429) supplemented with 10% FBS (Gibco, 10270106), 2 mM L-glutamine (Sigma-Aldrich, G7513), 50 μM ß-mercaptoethanol (Sigma-Aldrich, M3148), 10 mM HEPES (Sigma-Aldrich, H0887), 1 mM sodium pyruvate (Gibco, 11360039), 1X nonessential amino acids (Gibco, 11140035), 100 units/ml penicillin and 100 μg/ml streptomycin (Sigma-Aldrich, P0781)), followed by incubation on ice for 5 min. After three washing steps with 1 ml complete B cell medium (centrifugation: 2000 rpm for 2 min at 24°C; Eppendorf centrifuge 5424R), labeled cells were cultured in 500 μl B cell medium per well in 24-well plates. .. Following 48 h of culture, cells were gently resuspended, transferred to a 96-U-well plate, and stained (see section Cell surface staining ) with 30 μl of the following fluorochrome-conjugated anti-mouse antibodies diluted in FACS-B: αIgD-PerCP/Cy5.5 (1:200, Biolegend, 405710), αIgM-APC (1:1000, Jackson ImmunoResearch, 115-607-020), αCD19-BV711 (1:400, Biolegend, 115555), and αB220-BV785 (1:400, Biolegend, 103246).
Incubation:Article Title: TET2 loss promotes premalignant survival and clonal selection in MYC-driven B cell lymphoma
Article Snippet: .. Labeling was stopped by adding 4 volumes of cold complete B cell medium (DMEM medium (Sigma-Aldrich, D6429) supplemented with 10% FBS (Gibco, 10270106), 2 mM L-glutamine (Sigma-Aldrich, G7513), 50 μM ß-mercaptoethanol (Sigma-Aldrich, M3148), 10 mM HEPES (Sigma-Aldrich, H0887), 1 mM sodium pyruvate (Gibco, 11360039), 1X nonessential amino acids (Gibco, 11140035), 100 units/ml penicillin and 100 μg/ml streptomycin (Sigma-Aldrich, P0781)), followed by incubation on ice for 5 min. After three washing steps with 1 ml complete B cell medium (centrifugation: 2000 rpm for 2 min at 24°C; Eppendorf centrifuge 5424R), labeled cells were cultured in 500 μl B cell medium per well in 24-well plates. .. Following 48 h of culture, cells were gently resuspended, transferred to a 96-U-well plate, and stained (see section Cell surface staining ) with 30 μl of the following fluorochrome-conjugated anti-mouse antibodies diluted in FACS-B: αIgD-PerCP/Cy5.5 (1:200, Biolegend, 405710), αIgM-APC (1:1000, Jackson ImmunoResearch, 115-607-020), αCD19-BV711 (1:400, Biolegend, 115555), and αB220-BV785 (1:400, Biolegend, 103246).
Centrifugation:Article Title: TET2 loss promotes premalignant survival and clonal selection in MYC-driven B cell lymphoma
Article Snippet: .. Labeling was stopped by adding 4 volumes of cold complete B cell medium (DMEM medium (Sigma-Aldrich, D6429) supplemented with 10% FBS (Gibco, 10270106), 2 mM L-glutamine (Sigma-Aldrich, G7513), 50 μM ß-mercaptoethanol (Sigma-Aldrich, M3148), 10 mM HEPES (Sigma-Aldrich, H0887), 1 mM sodium pyruvate (Gibco, 11360039), 1X nonessential amino acids (Gibco, 11140035), 100 units/ml penicillin and 100 μg/ml streptomycin (Sigma-Aldrich, P0781)), followed by incubation on ice for 5 min. After three washing steps with 1 ml complete B cell medium (centrifugation: 2000 rpm for 2 min at 24°C; Eppendorf centrifuge 5424R), labeled cells were cultured in 500 μl B cell medium per well in 24-well plates. .. Following 48 h of culture, cells were gently resuspended, transferred to a 96-U-well plate, and stained (see section Cell surface staining ) with 30 μl of the following fluorochrome-conjugated anti-mouse antibodies diluted in FACS-B: αIgD-PerCP/Cy5.5 (1:200, Biolegend, 405710), αIgM-APC (1:1000, Jackson ImmunoResearch, 115-607-020), αCD19-BV711 (1:400, Biolegend, 115555), and αB220-BV785 (1:400, Biolegend, 103246).
Cell Culture:Article Title: TET2 loss promotes premalignant survival and clonal selection in MYC-driven B cell lymphoma
Article Snippet: .. Labeling was stopped by adding 4 volumes of cold complete B cell medium (DMEM medium (Sigma-Aldrich, D6429) supplemented with 10% FBS (Gibco, 10270106), 2 mM L-glutamine (Sigma-Aldrich, G7513), 50 μM ß-mercaptoethanol (Sigma-Aldrich, M3148), 10 mM HEPES (Sigma-Aldrich, H0887), 1 mM sodium pyruvate (Gibco, 11360039), 1X nonessential amino acids (Gibco, 11140035), 100 units/ml penicillin and 100 μg/ml streptomycin (Sigma-Aldrich, P0781)), followed by incubation on ice for 5 min. After three washing steps with 1 ml complete B cell medium (centrifugation: 2000 rpm for 2 min at 24°C; Eppendorf centrifuge 5424R), labeled cells were cultured in 500 μl B cell medium per well in 24-well plates. .. Following 48 h of culture, cells were gently resuspended, transferred to a 96-U-well plate, and stained (see section Cell surface staining ) with 30 μl of the following fluorochrome-conjugated anti-mouse antibodies diluted in FACS-B: αIgD-PerCP/Cy5.5 (1:200, Biolegend, 405710), αIgM-APC (1:1000, Jackson ImmunoResearch, 115-607-020), αCD19-BV711 (1:400, Biolegend, 115555), and αB220-BV785 (1:400, Biolegend, 103246).
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![<t>Pre-B</t> colony formation in mice post 5 day in vivo cumulative exposure to DBC Number of CFU-B colonies per million BM cells [a] or per set of femurs [b] in mice following 5 day in vivo exposure to DBC via oral voluntary pill ingestion, 10 days post plating in <t>mouse</t> <t>methylcellulose</t> media for pre-B cell selection. Data shown are Means ± SD. *Significantly different compared to control (p< 0.05).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_0818/pmc04470818/pmc04470818__nihms669765f1.jpg)
![In vitro exposure of mice BM cells to As+3 ± DBC or DBC-diol. Number of CFU-B colonies per million cells in untreated mice BM cells post 10 day plating in pre-B mouse methylcellulose media containing As+3 ± DBC or DBC-diol [a], or containing MMA+3 ± DBC or DBC-diol [b]. Data shown are Means ± SD. #Significantly different compared to effect of DBC or As+3 given as individual treatments at the selected dose. *Significantly different compared to control (p<0.05).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_0818/pmc04470818/pmc04470818__nihms669765f3.jpg)