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. 2015 Apr 2;6(11):8709–8721. doi: 10.18632/oncotarget.3325

Figure 6. Overexpression of CD44ICD increases the transcriptional activation of the stemness factors, Sox2, and Oct4.

Figure 6

(A) The transcriptional activation was measured with a reporter assay in wild-type (diagonal bars) and CD44KD MCF-7 (solid bars) cells. Cells were transfected with a Sox2 and Oct4 reporter vector alone or co-transfected with CD44 siRNA, CD44 or CD44ICD expression vectors. Following transfection, the cells were incubated for 12 hr and a vehicle (−) or 5 μM of GSI (+) were added. The cells were incubated for an additional 24 hr. The transcriptional activity was measured by luciferase activity described in “materials and methods”. (B) The expression of CD44ICD or of the truncated mutant constructs, ICD_ΔN17 and ICD_ΔC19 were detected with a western blot. (C) MCF-7 cells were transfected with the reporter vector alone or co-transfected with CD44-ICD or CD44-ICD_ΔN17 expression vectors for 36 hr. The luciferase activity was measured as described in “materials methods”. (D) MCF-7 cells were transfected with the reporter vector alone or co-transfected with CD44ICD or CD44ICD_ΔC19 expression vectors for 36 hr. The luciferase activity was measured. The data are presented as the mean ± SD (n = 3). Significant differences are indicated by an asterisk (*p < 0.05), and the p values were calculated using the Student's t test.