Cells were seeded into tissue culture flasks to give approximately 50% confluence, allowed to attach for 2–3 h, and then treated for a period of 48 h with either tamoxifen or fulvestrant in combination with afatinib. Each drug was administered at three dose levels, giving a fixed ratio (i.e., afatinib: 2.5 µM, 5.0 µM, and 10 µM; tamoxifen: 5 µM, 10 µM, and 20 µM; fulvestrant: 0.5 µM, 1.0 µM, and 2.0 µM). An annexin V–fluoroscein isothiocyanate (FITC)-conjugated apoptosis detection kit incorporating propidium iodide (PI) was used as described by the manufacturer’s protocols (Merck, Middlesex, UK). Harvesting of the cells included collection of attached cells following trypsinization as well as floating cells, which were combined in the sample for processing. Analysis by flow cytometry used the FL1 (FITC) and FL3 (PI) laser lines, and each sample was assessed using a collection of 10,000 events. For isobologram analysis, the Calcusyn program (version 211) was used (Biosoft®). Analyses were set up on three separate occasions.
Calcusyn program version 2
Manufactured by Biosoft
Sourced in United Kingdom
CalcuSyn program version 2.11 is a software application designed for data analysis and mathematical modeling. It provides core functions for dose-effect analysis and combination index calculation.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using calcusyn program version 2
1
Synergistic Apoptosis Induction by Targeted Therapies
2
Synergistic Drug Interaction Analysis
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Combination Index (CI) and isobologram were calculated by CalcuSyn program version 2.11 (Biosoft, Great Shelford, Cambridge, UK), and the effect of drug interactions was quantitatively assessed. CI values less than 1.0, 1.0, and greater than 1.0 demonstrate synergism, additivity, and antagonism, respectively. The isobologram is formed by plotting the doses of each drug required for 50% inhibition (ED 50 ) on the xand y-axis, and connecting them to draw a line segment, which is ED 50 isobologram. Combination data points that below, fall on, and above the line segment reveal synergism, additivity, and antagonism, respectively. All experiments were performed in triplicate.
3
Synergistic Drug Interaction Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Combination index (CI) and isobologram were calculated by CalcuSyn program version 2.11 (Biosoft, Great Shelford, Cambridge, UK), and the effect of drug interactions was quantitatively determined. CI values less than 1.0, 1.0, and greater than 1.0 demonstrate synergism, additivity, and antagonism, respectively. The isobologram is formed by plotting the doses of each drug required for 50% inhibition (ED 50 ) on the xand y-axes and connecting them to draw a line segment, which is ED 50 isobologram. Combination data points that fall on, below, and above the line segment indicate additivity, synergism, and antagonism, respectively. All experiments were performed in triplicate.
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