Fig. 6.

ALM201 alone or in combination with tamoxifen delays tumour recurrence in vivo which correlates with reduced number of mammospheres ex vivo. a Established MCF-7 xenografts (100–150 mm³) were treated with vehicle control (n = 5), tamoxifen (n = 4) or ALM201 (n = 4) alone or in combination (n = 4) for 21 days. Mammosphere formation was assessed ex vivo following excision and disaggregation of established MCF-7 xenografts; n ≥ 3 replicates per mouse. b Tumour cells from the treated xenografts were re-implanted into secondary mice and tumour occurrence was monitored twice a week and time to tumour initiation calculated (vehicle control, n = 13; tamoxifen (Tam), n = 14; ALM201, n = 5; tamoxifen + ALM201 (Tam + ALM201), n = 5). c Mammosphere formation following excision and disaggregation of established MCF-7 xenografts from the second generation mice without any further treatment in vivo (control, n = 6; Tam, n = 4; ALM201, n = 2; Tam + ALM201, n = 3); n ≥ 3 replicates per mouse. d Real-time qPCR analysis of DLL4 in MCF-7 xenografts treated with tamoxifen and ALM201 in vivo (n = 2). The difference in gene expression was presented as a fold change relative to the expression of the housekeeping genes, GAPDH and ß -Actin. Data points are mean ± SEM. n ≥ 3. * p < 0.05, ** p < 0.01, *** p < 0.001 (one-way ANOVA with post-hoc Dunnett’s multiple comparisons test)