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. 2021 Aug 24;4:1001. doi: 10.1038/s42003-021-02526-y

Fig. 4. Immunofluorescence analysis of pharmacodynamic biomarkers in SW620 tumour spheroids following different treatment schedules in microfluidic Ibidi chip.

Fig. 4

Spheroids were treated in the chip with SN38 and AZD0156 at different schedules using the microfluidic setup. Spheroids were recovered from the chip at day 7 and DNA double strand break damage was assessed via the presence of γH2AX. Scalebar = 100 µm. a Representative images of nuclei (blue/Hoechst 33342) and γH2AX (green), b quantification of γH2AX positive cells. (A-SN38, B-SN38 + AZD0156 3/7 with 24 h gap, C-SN38 + AZD0156 3/7 with 72 h gap, D-SN38 + AZD0156 1/7, E-SN38 + AZD0156 7/7, F-control). Cleaved caspase 3 (CC-3) was used to quantified apoptotic cell death (c), and Ki67 to measure the effect on proliferation (d). N ≥ 5 per condition from two independent experiments. Box-plots show median (centre line); box limits are 25th to 75th percentile; whiskers represent min and max values. Statistical analysis was carried out using 1-way ANOVA, Tukey’s multiple comparisons, CI = 95%, ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.1.