Figure 6. SRF–YAP–IL6 signalling is required for CSC formation.

(a–c) YAP/TAZ knockdown decreases IL6 expression and CSC formation. MCF-10A cells were transformed with a constitutively active form of H-Ras (HRas^G12V) and then transduced with shRNA against the indicated genes. (a) Western blot and qRT–PCR analyses (n=2 replicates), (b) representative FACS plot and statistical analyses of CD44 and CD24 antigen expression (n=3 experiments) and (c) statistical analyses of the number of mammospheres formed by MCF-10A cells expressing HRasG12V together with shRNA against YAP and/or TAZ (n=3 experiments). (d,e) MCF-10A H-Ras^G12V cells were infected with shRNA against SRF. (d) Western blot and qRT–PCR analyses (n=2 replicates) and (e) statistical analyses of the number of mammospheres formed by cells infected with the indicated viruses (n=3 experiments). (f,g) MCF-10A H-Ras^G12V cells infected with shRNA against YAP/TAZ or SRF were (f) infected with IL6-expressing retrovirus or (g) were treated with recombinant IL6 (r-hIL6; 5 ng ml⁻¹). Statistical analyses of the number of mammospheres formed by cells infected with the indicated viruses (n=3 experiments). (h) Limiting dilution xenograft assays of cells generated in f. Data are presented as means±s.e.m. (*P<0.05, **P<0.01, ***P<0.001, Student's t-test used in all analyses).