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