Twist directly upregulates ROR1 by occupying its promoter. (A) The indicated cells overexpressing empty vector or Twist were transfected with ROR1 promoter linked to luciferase, and, after 48 h, luciferase activity was assayed. (B) Illustration of the ROR1 promoter region with the indicated Twist-binding capability. "+" indicates binding and "-" indicates no binding based on the results shown in (C). (C) MDA-MB-231 cells were analyzed by ChIP using anti-Twist antibody. (D) Schematic illustration of wild-type ROR1 promoter and its mutants. (E) ROR1 promoters (wild-type or mutant) linked to luciferase were transfected into 293T cells, and, after 48 h, luciferase activity was assayed. (F) The indicated cells overexpressing empty vector or Twist were transfected with ROR1 promoter (wild-type or mutant) linked to luciferase, and, after 48 h, luciferase activity was assayed. (G, I) The indicated cells stably transfected with control or Twist shRNA were used to analyze the mRNA level of the indicated molecules by qRT-PCR. (H, J) The indicated cells stably transfected with control or Twist shRNA were used to analyze the protein levels by Western blot.