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. 2019 Apr 22;16:721–732. doi: 10.1016/j.omtn.2019.04.013

Figure 6.

Figure 6

Self-Renewal and Tumorigenic Potential of LCSCs Were Repressed by miR-1305 through the UBE2T-Mediated Akt-Signaling Pathway

(A) The gray value of Akt and GSK3β and their phosphorylation protein bands in LCSCs measured by western blot analysis. (B) The protein expressions of Akt and GSK3β and the extents of their phosphorylation in LCSCs, measured by western blot analysis. (C) The sphere formation assay in LCSCs treated with DMSO or LY294002 in the presence of miR-1305 mimic or UBE2T over-expression (100×). (D) The quantitative analysis of sphere formation ability in (C). (E) The soft agar colony formation assay in LCSCs treated with DMSO or LY294002 in the presence of miR-1305 mimic or UBE2T over-expression. (F) The quantitative analysis of soft agar colony formation in (E). (G) The proliferative ability of LCSCs that overexpressed miR-1305 or UBE2T and were treated with DMSO or LY294002, evaluated by CCK-8 assay. (H) Naked eye observation of tumor in nude mice treated with DMSO or LY294002 in the presence of miR-1305 mimic or UBE2T overexpression. (I) The tumor volume curve in nude mice treated with DMSO or LY294002 in the presence of miR-1305 mimic or UBE2T overexpression, measured by a Vernier caliper. (J) The tumor weight in nude mice in the presence of miR-1305 mimic or UBE2T overexpression with DMSO or LY294002 treatment. (K) The pathological characteristics of tumor in nude mice injected with miR-1305 mimic or UBE2T overexpression with DMSO or LY294002 treatment, observed with H&E staining (400×). miR-1305 scramble + DMSO group, LCSCs or nude mice transfected with miR-1305 mimic, then treated with DMSO; miR-1305 mimic + LY294002 group, LCSCs or nude mice transfected with miR-1305 mimic, then treated with LY294002; UBE2T + DMSO group, LCSCs or nude mice transfected with UBE2T overexpression, then treated with DMSO; UBE2T + LY294002 group, LCSCs or nude mice transfected with UBE2T overexpression, then treated with LY294002. *p < 0.05 versus LCSCs or nude mice transfected with miR-1305 mimic, then treated with DMSO; #p < 0.05 versus LCSCs or nude mice transfected with UBE2T overexpression, then treated with DMSO. Statistical data were described as mean ± SD. Data were analyzed by one-way ANOVA, and data in (G) and (I) were compared by repeated-measures ANOVA. The experiment was repeated 3 times independently (n = 6).