Figure 2: LncBRM is required for the self-renewal maintenance of liver CSCs.

(a) LncBRM-silenced cells from HCC samples were established and examined by northern blotting (left panel), followed by sphere formation assays. Representative sphere images were shown in the middle panel and statistic ratios were shown in the right panel. (b) Declined self-renewal capacities of lncBRM-depleted cells were detected by serial sphere formation assays. (c) LncBRM-depleted and control cells (1 × 10⁶) were injected into BALB/c nude mice, followed by measurement of tumour volumes. n=5 for each group. (d) LncBRM-silenced primary cells were established with pSiCoR lentivirus and used for subcutaneous injection. Xenograft tumours were used for serial tumour implantation. n=5 for each group. (e) LncBrm silenced and shCtrl cells (10, 10², 10³, 10⁴ and 10⁵) were subcutaneously injected into BALB/c nude mice for observation of tumour growth. Tumour-free mice ratios and tumour sizes are shown. (f) LncBRM-overexpressing primary cells were established and used for sphere formation assays. (g) LncBRM-overexpressing cells were examined by serial sphere formation assays. (h) LncBRM-overexpressing cells were injected into BALB/c nude mice for observation of tumour growth and measurement of tumour volume every 4 days. n=5 for each group. Scale bars, 500 μm (a,f). Data are shown as means±s.d. Two tailed Student’s t-test was used for statistical analysis; *P<0.05, **P<0.01 and ***P<0.001. Data are representative of four independent experiments.