il-3 culture supplement, mouse corning 354040  (Corning Life Sciences)

Journal: Oncotarget

Article Title: Paradoxical counteraction by imatinib against cell death in myeloid progenitor 32D cells expressing p210BCR-ABL

doi: 10.18632/oncotarget.25849

Figure Lengend Snippet: (A) 32D/TetOff-p210 cells were Tet-supplied or depleted and then cultured for 48 h in the presence or absence of imatinib (1 μM). WCL were subjected to immunoblotting. The values of relative band intensity versus the Tet (+) control were shown below each panel. Data are representative of three independent experiments. (B) 32D/TetOff-p210 cells or parental 32D cells were Tet-supplied or depleted and then cultured with IL-3 supplement for 48 h in the presence or absence of imatinib (1 μM). Cell viability was determined by WST-8 assay. * P < 0.01, ** P < 0.001. Data are shown as mean ± SEM ( n = 6) and are representative of three independent experiments. NS indicates no significant difference. (C) 32D/TetOff-p210 cells were Tet-supplied or depleted and then cultured for 96 h in the presence or absence of imatinib (1 μM). Cells were double-stained with annexin V and PI and analyzed by flow cytometry. The proportion of cell population with the representative data is shown in each panel. Four independent double-staining experiments were performed and statistical analysis was executed. * P < 0.01, ** P < 0.001. Data are shown as mean ± SEM ( n = 4).

Article Snippet: Mouse myeloid progenitor 32D cells were pharchased from RIKEN BioResouce Centaer Cell Bank (RBRC-RCB1145; Tsukuba, Ibaraki, Japan) and cultured in RPMI1640 supplemented with 10%(v/v) FBS and 10%(v/v) IL-3 culture supplement (Corning, NY, USA), defined as the 32D growth media, at 37°C under 5% CO2 in air.

Techniques: Cell Culture, Western Blot, Control, Staining, Flow Cytometry, Double Staining

Journal: Oncotarget

Article Title: Paradoxical counteraction by imatinib against cell death in myeloid progenitor 32D cells expressing p210BCR-ABL

doi: 10.18632/oncotarget.25849

Figure Lengend Snippet: (A) 32D/TetOff-p210 cells were Tet-depleted and then cultured for 48 h. WCL were subjected to immunoblotting. Bands were visualized by probing with antibodies against caspase-1 p10, cleaved caspase-3 or cleaved PARP. (B) 32D/TetOff-p210 cells were Tet-depleted or supplied and then cultured in the presence or absence of imatinib (1 μM) for 96 h and then incubated with FLICA 660 active caspase-1 or caspase-3 detection probe and analyzed by flow cytometry. The merged histogram (solid line) with non-stain control (dashed line) is shown in each panel. The proportion of FLICA-positive cell population max is shown in each panel. Three independent FLICA caspase-1/3 assays were performed, and statistical analysis was executed, as shown in lower graphs. * P < 0.05, ** P < 0.01. Data are shown as mean ± SEM ( n = 3).

Article Snippet: Mouse myeloid progenitor 32D cells were pharchased from RIKEN BioResouce Centaer Cell Bank (RBRC-RCB1145; Tsukuba, Ibaraki, Japan) and cultured in RPMI1640 supplemented with 10%(v/v) FBS and 10%(v/v) IL-3 culture supplement (Corning, NY, USA), defined as the 32D growth media, at 37°C under 5% CO2 in air.

Techniques: Cell Culture, Western Blot, Incubation, Flow Cytometry, Staining, Control

Journal: Oncotarget

Article Title: Paradoxical counteraction by imatinib against cell death in myeloid progenitor 32D cells expressing p210BCR-ABL

doi: 10.18632/oncotarget.25849

Figure Lengend Snippet: 32D/TetOff-p210 cells or parental 32D cells were Tet-supplied or depleted and then cultured with IL-3 supplement for 96 h to collect the culture supernatant and WCL. (A) ELISA was performed to determine concentration of IL-1β and TNF-α in the culture supernatant. * P < 0.001, # P < 0.05. Data are shown as mean ± SEM ( n = 3) and are representative of three independent experiments. ND indicates not detected (under the detection limit). NS indicates no significant difference. (B) ELISA was performed to determine concentration of S100A8/A9 in the culture supernatant. * P < 0.05. Data are shown as mean ± SEM ( n = 3). ND indicates not detected (under the detection limit). WCL were subjected to immunoblotting. Bands were visualized by probing with antibodies against S100A8, S100A9 or actin. The values of relative band intensity versus the Tet (+) control are shown under each panel. (C) ELISA was performed to determine concentration of S100A8/A9 in the plasma of 8-month-old WT ( n = 7) and BCR-ABL TG ( n = 9) mice. Data are shown as mean ± SEM. * P < 0.05. (D) Expressions of both S100a8 and S100a9 in the spleen of 8-month-old WT ( n = 5) and BCR-ABL TG ( n = 6) mice were determined by quantitative RT-PCR. Data are shown as mean ± SEM. * P < 0.005. Statistical significance was evaluated by Mann–Whitney U test.

Article Snippet: Mouse myeloid progenitor 32D cells were pharchased from RIKEN BioResouce Centaer Cell Bank (RBRC-RCB1145; Tsukuba, Ibaraki, Japan) and cultured in RPMI1640 supplemented with 10%(v/v) FBS and 10%(v/v) IL-3 culture supplement (Corning, NY, USA), defined as the 32D growth media, at 37°C under 5% CO2 in air.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Concentration Assay, Western Blot, Control, Clinical Proteomics, Quantitative RT-PCR, MANN-WHITNEY

Journal: Oncotarget

Article Title: Paradoxical counteraction by imatinib against cell death in myeloid progenitor 32D cells expressing p210BCR-ABL

doi: 10.18632/oncotarget.25849

Figure Lengend Snippet: 32D/TetOff-p210 cells were Tet-supplied or depleted and then cultured in the presence or absence of imatinib (1 μM) for 96 h. (A) Giemsa staining was performed. Cells with segmented nuclei (indicated by arrows) are shown in each panel. Scale bar, 10 μm. (B) Cells were triple-stained with anti-CD11b-BV421, anti-Ly6C-APC, and anti-Ly6G-PE, or isotype anti-rat IgG2b-BV421, anti-rat IgM-APC, and anti-rat IgG2a-PE. Stained cells were analyzed by flow cytometry. The obtained data were processed by selection of PI - and CD11b + cell population, and then the cell surface expression of Ly6C and Ly6G within the CD11b + cell population was analyzed. Numbers in the plots indicate the percentages of gated cells. (C) Three independent triple-staining experiments, as exemplified in panel B, were performed, and statistical analysis was executed. * P < 0.01. Data are shown as mean ± SEM ( n = 3).

Article Snippet: Mouse myeloid progenitor 32D cells were pharchased from RIKEN BioResouce Centaer Cell Bank (RBRC-RCB1145; Tsukuba, Ibaraki, Japan) and cultured in RPMI1640 supplemented with 10%(v/v) FBS and 10%(v/v) IL-3 culture supplement (Corning, NY, USA), defined as the 32D growth media, at 37°C under 5% CO2 in air.

Techniques: Cell Culture, Staining, Flow Cytometry, Selection, Expressing