Figure 6: LncBRM-mediated YAP1 signalling is required for self-renewal of liver CSCs and tumour propagation.

(a) YAP1 was activated (overexpression) or inactivated (Verteporfin treatment) in non-sphere cells or sphere cells, followed by measurement of self-renewal by serial sphere formation assays. Fourth passage spheres and sphere numbers were shown (upper two panels). Two weeks later, original spheres, Verteportin-treated spheres and YAP1-overexpressing spheres were collected for mRNA detection (lower panels). (b) YAP1 KO cells (YAP1KO) were used for sphere formation assays. (c) Wild-type (WT) and Yap1-deficient cells (10, 10², 10³, 10⁴ and 10⁵) were injected into BALB/c nude mice for measurement of tumour formation. Tumour-free mice and established tumours were shown. n=6 for each group. (d) 1 × 10⁶ YAP1 KO and control cells were injected into BALB/c nude mice for tumour growth. Tumour volume was calculated every 4 days. n=5 for each group. (e) HCC primary tumour cells were treated with YM155 and Siomycin A for sphere formation assay. (f) YAP1 was rescued in lncBRM-depleted cells using pBPLV lentivirus and followed by sphere formation assay. Scale bars, 500 μm (a,b,e,f). Data are shown as means±s.d.; *P<0.05, **P<0.01 and ***P<0.001. Data are representative of four independent experiments.