Figure 4.
p120-catenin plays a crucial role for bridging TIM-3/Gal-9 signaling to the canonical Wnt pathway in AML cells. (A) Cellular growth of KASUMI-3 cells transfected with scrambled control and shRNA-mediated CTNND1 KD vectors. Two kinds of shRNA (shCTNND1-1: target sites in the 3' untranslated region, and shCTNND1-2: target sites in the coding sequence) were used. Data are presented as mean ± SEM, ∗P < .05 and ∗∗P < .01 vs scrambled control. (B) WB analysis of the canonical Wnt pathway–related protein using cell lysates from KASUMI-3 cells transfected with scrambled control and 2 kinds of shRNA-targeting CTNND1. (C) WB analysis of the phosphorylation status of p120-catenin at Tyr228 induced by stimulation with Gal-9 (AML#7). (D) Quantification of the levels of phosphorylated p120-catenin at Tyr228 by Gal-9 stimulation in KASUMI-3 cells. Results of 4 independent experiments (AML#1, #7, #9, and #16) are summarized. Data are presented as mean ± SEM, ∗P < .05 vs nonstimulated control. (E) WB analysis of the inhibitory effect of A-419259 (0, 1, or 10 nM) on Gal-9 stimulation–induced phosphorylation of p120-catenin using a primary AML sample (AML#9). (F) Quantification of the levels of phosphorylated p120-catenin induced by Gal-9 stimulation in the presence of each concentration of A-419259 (0, 1, and 10 nM). Data are presented as mean ± SEM, ∗P < .05 and ∗∗P < .01. Results from 4 independent AML experiments (AML#6, #9, #14, and #17) are summarized. (G) Immunoblotting analysis with total p120-catenin and phosphorylated p120-catenin (Tyr228) antibodies of total cell lysates (left), and immunoprecipitated lysates (right). Cells were stimulated with Gal-9 for 0, 5, and 10 minutes, and subsequently lysed and subjected to IP with an anti-HCK antibody or normal rabbit IgG. Representative results out of 3 independent experiments are shown here.

p120-catenin plays a crucial role for bridging TIM-3/Gal-9 signaling to the canonical Wnt pathway in AML cells. (A) Cellular growth of KASUMI-3 cells transfected with scrambled control and shRNA-mediated CTNND1 KD vectors. Two kinds of shRNA (shCTNND1-1: target sites in the 3' untranslated region, and shCTNND1-2: target sites in the coding sequence) were used. Data are presented as mean ± SEM, ∗P < .05 and ∗∗P < .01 vs scrambled control. (B) WB analysis of the canonical Wnt pathway–related protein using cell lysates from KASUMI-3 cells transfected with scrambled control and 2 kinds of shRNA-targeting CTNND1. (C) WB analysis of the phosphorylation status of p120-catenin at Tyr228 induced by stimulation with Gal-9 (AML#7). (D) Quantification of the levels of phosphorylated p120-catenin at Tyr228 by Gal-9 stimulation in KASUMI-3 cells. Results of 4 independent experiments (AML#1, #7, #9, and #16) are summarized. Data are presented as mean ± SEM, ∗P < .05 vs nonstimulated control. (E) WB analysis of the inhibitory effect of A-419259 (0, 1, or 10 nM) on Gal-9 stimulation–induced phosphorylation of p120-catenin using a primary AML sample (AML#9). (F) Quantification of the levels of phosphorylated p120-catenin induced by Gal-9 stimulation in the presence of each concentration of A-419259 (0, 1, and 10 nM). Data are presented as mean ± SEM, ∗P < .05 and ∗∗P < .01. Results from 4 independent AML experiments (AML#6, #9, #14, and #17) are summarized. (G) Immunoblotting analysis with total p120-catenin and phosphorylated p120-catenin (Tyr228) antibodies of total cell lysates (left), and immunoprecipitated lysates (right). Cells were stimulated with Gal-9 for 0, 5, and 10 minutes, and subsequently lysed and subjected to IP with an anti-HCK antibody or normal rabbit IgG. Representative results out of 3 independent experiments are shown here.

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