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13 protocols using cell titer glo luminescence cell viability kit

1

Synergistic Effects of SSZ, NAC, and CDDP in MBT-2V Cells

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MBT‐2V cells were seeded on 96‐well black plates in a volume of 4 × 103 cells/100 μL/well culture medium and incubated at 37°C for 24 hours under 5% CO2 in a humidified incubator. Cells were treated with various concentrations of SSZ with or without NAC (3 μmol/L) or with or without CDDP (10 μmol/L) for 48 hours. The number of viable cells was evaluated with a Cell Titer‐Glo luminescence cell viability kit (Promega, Madison, WI, USA). The luminescence signal was detected with an Enspire 2300 (PerkinElmer, Waltham, MA, USA). Data were expressed as the percentage of viable cells relative to that of the control. The experiment was carried out in triplicate, and data were expressed as the mean ± standard error (SE) of relative cell viability. A combination index (CI) analysis was also used to measure of the extent of the drug interaction quantitatively. A combination index (CI) of less than, equal to, and more than 1 indicates synergy, additivity, and antagonism, respectively.23
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2

Trolox Effect on CCA Cell Viability and Migration

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The KKU-213 CCA cell line with either transduced control or CD44shRNA (2x103 cells per well) was treated with Trolox for 24 h. The number of viable cells was evaluated with a Cell Titer-Glo luminescence cell viability kit (Promega, Madison, WI). The luminescence signal was detected on a SpectraMaxL microplate reader.
In addition, the CCA KKU-213 cell line with either transduced control or CD44shRNA (2x104 cells per well) with serum free DMEM with the absence or present of Trolox was placed on the upper layer of a cell permeable membrane and a solution containing 10% fetal bovine serum was placed below the cell permeable membrane for 24 h. Next, migrated cells were fixed with methanol and then stained with hematoxylin (Sigma-Aldrich). The number of migrated cells were counted as cells per field under a Microscope Axio Imager A2 (Carl-Zeiss, Oberkochen, Germany, located in Khon Kaen, Thailand).
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3

HER2 Mutant Cell Viability Assay

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BaF3 cells stably expressing HER2 mutants were washed twice with PBS and plated in 96-well plates (10,000 cells/well) in replicates of 12 in complete RPMI medium without IL-3. Cell viability was measured using the Cell Titer Glo Luminescence Cell Viability Kit (Promega), and plates were read on a Synergy 2 (Biotek Instruments) luminescence plate reader. Relative survival reported in cell survival studies is a ratio of relative luciferase activity (RLU) at day 4 over RLU measured at the time (day 0) the experiment was initiated.
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4

Evaluating CCA Cell Viability

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The number of viable cells was evaluated with a Cell Titer-Glo luminescence cell viability kit (Promega, Madison, WI, USA). Briefly, CCA cells (2x103 cells per well) were plated into 96-black well plates for 24 h. Cells were then treated with SSZ (0, 200, 400, 600, 800, 1000 μM) and CIS (0, 10, 20, 40, 60, 80, 100 μM) for 48 h, and GEM (0, 10, 20, 40, 60, 80, 100 μM) for 72 h. In addition, 300 μM of SSZ was used in combination with CIS or GEM. The luminescence signal was detected on a SpectraMaxL microplate reader. The experiments were done in triplicate.
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5

Cell Attachment on ECM Proteins

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B16-BL6 and MeWo cells (2.0 × 104 per well) in 100 μl of serum-free medium were transferred to 96-well flat-bottom plates coated with POSTN, POSTN-ΔC, COL-I, or FN as described above for the scratch wound assay. Noncoated wells served as controls. The cells were incubated for 90 min at 37°C, the wells were washed twice with PBS, and attached cells were assayed for each well with the use of a Cell Titer-Glo luminescence cell viability kit (Promega, Madison, WI). In some experiments, B16-BL6 cells were incubated with 10 μg/ml of anti–mouse integrin αv Ab or 10 μg/ml of rat IgG1 isotype control Ab for 2 h and cells were then transferred to 96-well flat-bottom plates. The resulting values are expressed as percentages relative to the control (100%).
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6

Cell Proliferation on ECM Coatings

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B16-BL6 and MeWo cells suspended in 100 μl of medium supplemented with 10% FBS were seeded at a density of 2.0 × 103 cells per well in 96-well flat-bottom plates coated with POSTN, COL-I, or FN as described above. Noncoated wells served as controls. The cells were cultured for 72 h, washed twice with PBS, and assayed for cell proliferation with the use of a Cell Titer-Glo luminescence cell viability kit (Promega). Data are expressed relative to the values of cells cultured for 12 h.
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7

Measuring Cell Viability via ATP

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Viability was assessed by measuring ATP levels following CBL0100 treatment using Promega Cell-Titer-Glo Luminescence cell viability kit (G7570), following the manufacturer's instructions. The luciferase signal was measured using the Cytation 5 imaging reader (BioTek).
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8

Cell Viability Assay for CCA Cells

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The number of viable cells was evaluated with a Cell Titer-Glo luminescence cell viability kit (Promega, Madison, WI). In short, CCA cell lines with either transduced control or CD44shRNA (2x103 cells per well) were plated into 96-black well plates for 24, 48, 72 and 96 h. The luminescence signal was detected on a SpectraMaxL microplate reader. instructions. Experiments were performed three times with three replicates per experiment.
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9

Cell Viability Assay in Ba/F3 Cells

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Ba/F3 pro‐B cells (DSMZ, Germany) were washed with PBS and resuspended in media (RPMI +10% FBS) without IL‐3, then plated at 10 000 cells/well in 96‐well plates. On day 0 and day 4 cell viability was measured using Cell Titer‐Glo Luminescence Cell Viability kit (Promega) and plates were read on a Synergy 2 luminescence plate reader (Biotek Instruments). The luminescence reading on day 4 divided by the reading on day 0 was used to express relative cell survival.
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10

HGF-Induced Growth Assay in NIH3T3 Cells

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NIH3T3 cells stably expressing wild-type or Asp153Tyr mutant MET were plated in complete medium. After 24 h, the medium was replaced with serum-free medium and cells were treated with the indicated concentration of recombinant HGF (R&D Systems). Cell growth was measured after 3 d with the CellTiter-Glo luminescence cell viability kit (Promega) as described previously98 (link). Student's t test (two-tailed) was used for statistical analyses to compare treatment groups with GraphPad Prism 5.00. A P value of <0.05 was considered statistically significant (*P < 0.05 and **P < 0.01).
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