Figure 3.

TELO2 is essential for the maintenance of β-catenin levels

(A) Knockdown of TELO2 with siRNAs reduced β-catenin/TCF-dependent transcriptional activation in HEK293 cells. The cells transfected with control or TELO2 siRNAs, and then transiently cotransfected with Super 8x TOPFlash (a firefly luciferase reporter plasmid) to assess the β-catenin/TCF-dependent transcriptional activity and with pRL-SV40 (a renilla luciferase reporter plasmid) to normalize the transfection efficiency. The cells were treated with 50 ng/mL Wnt3A for 18 h. Firefly and renilla luciferase activities were measured. Normalized relative luciferase activities were calculated by dividing firefly luciferase activities by those of renilla and indicated the level of transcriptional activation. Data are presented as the means ± SDs (n = 3 biological replicates). ∗∗∗p < 0.001, one-way ANOVA with Tukey’s test.

(B–D) Knockdown of TELO2 with siRNAs reduced the β-catenin levels in HEK293 and HT-29 cells. HEK293 (B and C) or HT-29 (D) cells were transfected with siRNAs for TELO2 for 72 h. The cell lysates were analyzed via western blotting with the indicated antibodies (B and D).

(C and D) Band intensities were quantified, normalized to the actin levels, and reported as values relative to those obtained from the cells transfected with a control siRNA. Data are presented as the means ± SDs (C, n = 3 biological replicates; D, n = 4 biological replicates). ∗∗p < 0.01, ∗∗∗p < 0.001, one-way ANOVA with Tukey’s test.