Figure 2
Figure 2. Deletion of the β-catenin binding site in p210 BCR/ABL diminishes the accumulation of p-β-catenin (Tyr654) and TCF/LEF-mediated transcription. (A) 293T cells were transiently transfected with p210 BCR/ABL (BCR/ABL), the ΔUBD mutant, or cognate vector. WCLs were collected and examined by western blot for expression of the constructs. Lysates were also used for IPs using a β-catenin antibody, and immunoprecipitates were examined by western blot using antibodies for β-catenin and p210 BCR/ABL. (B) 293T cells were transfected with p210 BCR/ABL in the presence or absence of the E1 inhibitor. WCLs were collected and examined by western blot for expression of p210 BCR/ABL (BCR/ABL), cyclin D2, or RhoA. Lysates were also used for IPs using a β-catenin antibody, and immunoprecipitates were examined by western blot using antibodies for β-catenin and BCR/ABL. (C) Ba/F3 cells were infected with retroviral particles that encode MSCV-bcr-abl/p210-IRES-gfp (BCR/ABL), MSCV-bcr-abl/p210(ΔUBD)-IRES-gfp (ΔUBD), or cognate vector. GFP+ cells were then sorted and cultured. Total cell lysates (T) were collected and examined by western blot for expression of the constructs and β-catenin. Nuclear (N) and cytosolic (C) lysates were prepared and examined by western blot as indicated. Quantified data for p-β-catenin (Tyr654) expression are shown on the right. (D) WCLs were also used for IPs using a BCR/ABL antibody, and immunoprecipitates were examined by western blot using antibodies for β-catenin and BCR/ABL as indicated. (E) TOP- and FOPflash plasmids were transfected into the stably infected Ba/F3 cells. Twenty-four hours after transfection, cells were analyzed for luciferase activity. (F) WCLs from stably infected Ba/F3 cells were examined by western blot for expression of cyclin D1 and c-Myc. All quantitative data shown are averages ± standard deviations of at least 3 independent experiments. P values were calculated by using a paired Student t test or a one-way ANOVA followed by Tukey tests. *,#P < .05; **,##P < .01; ***,###P < .001. *Indicates significance relative to vector; #indicates significance relative to p210 BCR/ABL.

Deletion of the β-catenin binding site in p210 BCR/ABL diminishes the accumulation of p-β-catenin (Tyr654) and TCF/LEF-mediated transcription. (A) 293T cells were transiently transfected with p210 BCR/ABL (BCR/ABL), the ΔUBD mutant, or cognate vector. WCLs were collected and examined by western blot for expression of the constructs. Lysates were also used for IPs using a β-catenin antibody, and immunoprecipitates were examined by western blot using antibodies for β-catenin and p210 BCR/ABL. (B) 293T cells were transfected with p210 BCR/ABL in the presence or absence of the E1 inhibitor. WCLs were collected and examined by western blot for expression of p210 BCR/ABL (BCR/ABL), cyclin D2, or RhoA. Lysates were also used for IPs using a β-catenin antibody, and immunoprecipitates were examined by western blot using antibodies for β-catenin and BCR/ABL. (C) Ba/F3 cells were infected with retroviral particles that encode MSCV-bcr-abl/p210-IRES-gfp (BCR/ABL), MSCV-bcr-abl/p210(ΔUBD)-IRES-gfp (ΔUBD), or cognate vector. GFP+ cells were then sorted and cultured. Total cell lysates (T) were collected and examined by western blot for expression of the constructs and β-catenin. Nuclear (N) and cytosolic (C) lysates were prepared and examined by western blot as indicated. Quantified data for p-β-catenin (Tyr654) expression are shown on the right. (D) WCLs were also used for IPs using a BCR/ABL antibody, and immunoprecipitates were examined by western blot using antibodies for β-catenin and BCR/ABL as indicated. (E) TOP- and FOPflash plasmids were transfected into the stably infected Ba/F3 cells. Twenty-four hours after transfection, cells were analyzed for luciferase activity. (F) WCLs from stably infected Ba/F3 cells were examined by western blot for expression of cyclin D1 and c-Myc. All quantitative data shown are averages ± standard deviations of at least 3 independent experiments. P values were calculated by using a paired Student t test or a one-way ANOVA followed by Tukey tests. *,#P < .05; **,##P < .01; ***,###P < .001. *Indicates significance relative to vector; #indicates significance relative to p210 BCR/ABL.

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