Figure 4: Treatment efficacy in 3D tumour spheroid model.

(a) 3D spheroid cultures of cancer cells grown on Matrigel. Top, micrographs showing AU565 and BT474 spheroids after 12 days with indicated treatment, for abbreviations see Fig. 2a. Bottom, size distribution of treated spheroids. Ten micrographs were analysed with Image J software (Mann–Whitney test). Scale bar, 50 μm. (b) Proliferation of treated 3D cultures of AU565, BT474, SKBR3 and HCC1419 in Matrigel were analysed by a modified XTT assay (*P value versus mock or as drawn, two-sided, unpaired Welch’s t-test). (c) AKT re-phosphorylation in 3D spheroids. BT474 cultures on Matrigel were treated by TZB or 6L1G and whole-cell extracts were analysed by immunoblots.