Xenbase Image ID: 168765

Figure 1. Constitutive TCF/LEF activity in hematopoietic tumor cells in the absence of β-catenin stabilization.(A) The indicated hematopoietic tumor cells were transduced with wild-type (Top) or mutant (Fop) TCF/LEF firefly luciferase reporter and a renilla luciferase virus, and the TCF/LEF transcriptional activity was calculated by dividing the TOP/renilla ratio by the FOP/renilla ratio. The mean values (± s.e.m., n>5) of at least two independent experiments are shown. (B) Lack of β-catenin stabilization in K562, Jurkat and Ramos cells, as measured by GST-E-cadherin pull-down. Ovarian PA1 cancer cells and breast immortalized AB5 cells were used as positive and negative control, respectively. (C) β-catenin or γ-catenin knockdown in Ramos cells did not affect TCF/LEF activity. Ramos cells containing Top or Fop luciferase reporter were infected with β-catenin or γ-catenin shRNA and the TCF/LEF reporter activity was measured (lower panel). β-catenin and γ-catenin down-regulation was assessed by immunoblot (upper panel). (D) Ramos and U937 cells containing the tet transactivator (tTA) with or without a lentiviral inducible vector containing DN-TCF4 fused to GFP were cultured for 24 hrs in the presence or the absence of doxycycline (dox), and the proportion of GFP positive cells was assessed by FACS analysis. (E) The cells in D were used in colony formation assay in the presence or the absence of dox. (F) Effects of DN-TCF4 induction in Ramos cells on the expression of LEF1 and Axin 2 by quantitative real-time PCR. (G) mRNA levels of Axin 2 and LEF1 in human primary hematopoietic tumors. For each sample, the qPCR values were normalized with those obtained for the negative control, U937 cells. The types of malignancy corresponding to each sample are listed in the table: MCL, mantle cell lymphoma; CLL, B-cell chronic lymphocytic leukemia; splMZL, splenic marginal zone lymphoma; FLN, follicular lymphoma; HLN, Hodgkin's lymphoma. Image published in: Grumolato L et al. (2013) Image reproduced on Xenbase with permission of the publisher and the copyright holder. Creative Commons Attribution license Larger Image Printer Friendly View

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