AKR1B1 overexpression highly correlates with BLBC. (A) Dot plots indicate AKR1B1 expression in different subtypes of breast cancer from microarray datasets (GSE25066 and NKI295). Comparisons were analyzed by one-way ANOVA. (B) Expression of AKR1B1 and Twist2 was analyzed by Western blotting in fresh frozen tumor samples from five cases of luminal and five cases of triple-negative breast cancer (other analyses of 20 cases of fresh tumor samples are shown in Fig. S1 B). (C) Dot plots indicate AKR1B1 expression in luminal and BLBC cell lines from four microarray datasets (GSE12777, GSE10890, GSE16732, and E-TABM-157). Comparisons were made using two-tailed Student’s t test. (D and E) Expression of AKR1B1 mRNA was examined by either semiquantitative RT-PCR (D) or quantitative real-time PCR (E) in a representative panel of breast cancer cell lines. Data are mean ± SD. *, P < 0.05 by Student’s t test. (F) Expression of AKR1B1, Twist2, E-cadherin, and other EMT markers in cells from D was analyzed by Western blotting. E-cad, E-cadherin; Lum, luminal; N-cad, N-cadherin.