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. 2021 Sep 21;11(19):9571–9586. doi: 10.7150/thno.61711

Figure 6.

Figure 6

Onvansertib effects in combination with reference treatments (cisplatin and radiotherapy) on sensitive and resistant CAL33 cell lines. (A) Cells were treated 72 h with onvansertib (25 nM) in combination with cisplatin (3 µM) and radiotherapy (8 Gy). Cell death was evaluated by flow cytometry (n = 3). Cells were stained with Propidium Iodure (PI) and Annexin V (AV). Histograms show AV+/PI- cells (early-apoptosis) and AV+/PI+ cells (late-apoptosis or another cell death). (B-C) Cells were treated with onvansertib (2.5 nM), cisplatin (1 µM) and radiotherapy (2 Gy). Quantification (B) and representative image of clonogenicity assay (C) (n = 3). (D-E) 3D cell culture assay (spheroid) was treated with the combination of the three treatments: onvansertib (25 nM and 50 nM), cisplatin (3 µM) and radiation (2 Gy) and evaluated after 6 days. Quantification (D) and representative image of invasion assay (E) (n = 2). Statistics were analyzed using ANOVA test: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 and unpaired Student's t test: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.