Fig. 3.

Wnt5a/b and β-catenin expression in carcinoma tissues from non-IBC and IBC patients and their correlations with AHR/CYP1B1 and patents clinical pathological properties. (A) Representatives of protein expression of Wnt5a/b and β-catenin in non-IBC and IBC carcinoma tissue lysates analyzed by SDS-PAGE, transferred into PVDF membranes showing high expression of Wnt5a/b and β-catenin in breast carcinoma tissues of IBC patients. (B) Bars represent the relative density values of protein bands assessed by ImageJ software and normalized against the loading control β-tubulin and showing significant expression of Wnt5a/b and β-catenin in IBC tissues (n = 28) compared to non-IBC tissues (n = 33). Data represent the mean ± SD, ^*P ≤ 0.05 and ^**P ≤ 0.001 as determined by Student’s t-test. (C and D) Scatter charts showing no correlation between AHR and CYP1B1 expression and Wnt5a/b and β-catenin expression in non-IBC carcinoma tissues. (E and F) Scatter charts showing a linear positive correlation between AHR/CYP1B1 expression and Wnt5a/b and β-catenin expression in IBC carcinoma tissues. (G and H) Bars represent expression of Wnt5a/b and β-catenin in non-IBC and IBC patients sub-grouped according to the tumour grade, both correlates with tumour grade in the non-IBC (G) and IBC (H) patient groups. (I and J) Bars represent the relative density values of Wnt5a/b and β-catenin proteins sub-grouped into negative and positive LVI, results showed no statistical correlation with LVI in non-IBC patients and a significant correlation with lymphovascular invasion in IBC patients. Data represent mean ± SD, ^*P ≤ 0.05 and ^**P ≤ 0.001 as determined by one-way ANOVA followed by Tukey’s multiple comparison test. (K – N) Scatter charts showing a weak correlation between the number of metastatic lymph nodes and the relative density values of Wnt5a/b (K) and β-catenin (L) proteins in non-IBC tissues and a linear positive correlation between the number of metastatic lymph nodes and the relative density values of Wnt5a/b (M) and β-catenin (N) proteins in IBC tissues. Correlation coefficient (r values) were calculated by Pearson’s correlation test. (O and P) Bars represent the fold change (RQ = 2^−ΔΔCt) of mRNA expression of Wnt5a in MDA-MB-231 (O) and SUM149 (P) cells after treatment with an AHR inhibitor. The results are representative of at least three independent experiments. Data represent the mean ± SD. ^*P ≤ 0.05 and ^**P ≤ 0.001 as determined by Student’s t test.