Evaluating APE1Ref-1 Inhibitor Efficacy

RP-B-01 and RP-B-02 cells were resuspended in normal growth media containing 3% Reduced Growth Factor Matrigel (BD Biosciences) at a cell density of 1,500 and 3,000 cells/well, respectively and plated in ultra-low adherence 96-well plates (Corning). After spheroid formation, APE1Ref-1 inhibitors were added on days 4, 8, and 12 as described in Arpin, et al (40 (link)). On Day 15, Alamar blue reagent (LifeTechnologies) was added to each well (10 μL/well) and all wells were incubated for 24 hours. IC₅₀ values were calculated for each compound and the linear regression model was employed to generate a line of best fit, wherein the percent survival equaled 50% (n=3-4).

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Fishel M.L., Xia H., McGeown J., McIlwain D.W., Elbanna M., Craft A.A., Kaimakliotis H.Z., Sandusky G.E., Zhang C., Pili R., Kelley M.R, & Jerde T.J. (2019). Anti-tumor activity and mechanistic characterization of APE1/Ref-1 inhibitors in bladder cancer. Molecular cancer therapeutics, 18(11), 1947-1960.

Publication 2019

Reduced Growth Factor Matrigel ultra-low adherence 96-well plates Alamar blue reagent

Alamar blue Cells Growth factor Inhibitors Matrigel Normal growth media

Corresponding Organization : Indiana University – Purdue University Indianapolis

Other organizations : University of Ulster, Indiana University Health, Purdue University West Lafayette

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Variable analysis

independent variables

APE1Ref-1 inhibitors

dependent variables

Cell survival (percent)
IC50 values

control variables

Normal growth media containing 3% Reduced Growth Factor Matrigel (BD Biosciences)
Cell density (1,500 cells/well for RP-B-01 and 3,000 cells/well for RP-B-02)
Culture in ultra-low adherence 96-well plates (Corning)
Spheroid formation prior to addition of APE1Ref-1 inhibitors

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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