Fig. 2

LEF1 is predominantly expressed in esophageal CSCs and promotes a CSC-like phenotype in ESCC cells in vitro. a, b qRT-PCR and western blotting analysis of magnetically sorted OV6⁺ adherent (a) and spheroid(b) subpopulations was performed to evaluate the relative mRNA and protein expression levels, respectively, of LEF1 in ECA109 and TE1 cells. Data are shown as the mean ± SD, *P < 0.05, **P < 0.01. c qRT-PCR analysis of LEF1 expression of ECA109 and TE1 cells in different culture conditions. Data are shown as the mean ± SD, **P < 0.01. d qRT-PCR analysis was performed for LEF1 and the stem cell-associated genes in magnetically sorted LV-LEF1 OV6⁺ or LV-shLEF1 OV6⁺ cells of two ESCC cell lines. Data are shown as the mean ± SD, *P < 0.05, **P < 0.01. e, f The protein level of LEF1 in the LV-LEF1 group and LV-shLEF1 group was shown in ECA109 and TE1 cells. g Tumor spheroid formation assay indicated that LV-LEF1 OV6⁺ cells were able to generate an increased number and size of primary and secondary spheroids in ECA109 and TE1 cell lines, whereas LEF1-silenced OV6⁺ cells exhibited the opposite effect (scale bar = 100um). Data are shown as the mean ± SD *P < 0.05, **P < 0.01. All experiments were performed in triplicate