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. 2018 Jan 9;7:e29747. doi: 10.7554/eLife.29747

Figure 3. Olaparib sensitivity after depletion of EWS-FLI1 from A673 cells.

A673 Ewing’s sarcoma cells transiently transfected with negative control siRNA (siCT) or an siRNA targeting the EWS-FLI1 translocation (siEF1). (A) Cell viability assays were performed with the indicated doses of olaparib, or an equivalent volume of vehicle control (DMSO). After 72 hr of treatment cell viability was determined. Relative viability was calculated as a percentage of untreated cells. Means reported and error bars represent SD from three independent biological repeats. (B) siRNA-mediated depletion of EWS-FLI1 was determined after 72 hr treatment with vehicle control (DMSO) or olaparib. Relative expression levels of EWS-FLI1 expression normalized to RPLP0 (ribosomal protein lateral stalk subunit P0) was determined by qRT-PCR. Expression level of siCT cells treated with DMSO was assigned a value of 100. Means reported and error bars represent SD from three independent biological repeats. Two-way ANOVA main effect for siRNA (siCT or siEF1); F(1,12) = 96, p=4.46×10−7. Pairwise contrast between DMSO treated cells transfected with siCT or siEF1; t(12) = 7.32, uncorrected p=9.17×10−6 with a priori alpha level = 0.025; (Bonferroni corrected p=1.83×10−5). Pairwise contrast between olaparib treated cells transfected with siCT or siEF1; t(12) = 6.53, uncorrected p=2.80×10−5 with a priori alpha level = 0.025; (Bonferroni corrected p=5.60×10−5). Additional details for this experiment can be found at https://osf.io/2w22x/.

Figure 3.

Figure 3—figure supplement 1. Cell viability assays for each biological repeat.

Figure 3—figure supplement 1.

This is the same experiment as in Figure 3. The line graph of each biological repeat is plotted individually. Means reported and error bars represent SD from three technical replicates for each measurement. Additional details for this experiment can be found at https://osf.io/2w22x/.