Additional file 1 of High B3GALT5 expression confers poor clinical outcome and contributes to tumor progression and metastasis in breast cancer

Additional file 1: Figure S1. Clinical and pathological characteristics of 202 breast cancer patients. Figure S2. Correlation between B3GATL5 expression levels and stage or grade of breast cancers. Tumor (a and b) or adjacent non-tumor (c and d) tissues from patients with grade I, II, and III (a and c) or stage I, II, and III-IV (b and d) were analyzed for expression of B3GALT5 mRNA by using qRT-PCR. Statistical analysis was performed One-way ANOVA analysis. Figure S3. Expression levels of B3GALT5 in tumor of breast cancer tissues. The GSE1456 dataset was used to plot the survival curve comparing the patient with high (black) and low (gray) expression of B3GALT5. Figure S4. Higher expression of B3GALT5 in breast cancer tissue correlates with poor clinical outcome (RFS). Kaplan-Meier plots of relapse-free survival (RFS) comparing the 202 breast cancer patients with high (black) and low (gray) expression of B3GALT5 in tumor (a-c) and adjacent non-tumor parts (d-f) of patients with stage I-II (a and d), and stage I-II & Luminal A (b and e), and stage I-II & Luminal B (c and f) breast cancer were analyzed. Figure S5. Higher expression of B3GALT5 in breast cancer tissue correlates with poor clinical outcome (OS). Kaplan-Meier plots of overall survival (OS) comparing the 202 breast cancer patients with high (black) and low (gray) expression of B3GALT5 in tumor (a-c) and adjacent non-tumor parts (d-f) of patients with stage I-II (a and d), and stage I-II & Luminal A (b and e), and stage I-II & Luminal B (c and f) breast cancer were analyzed. Figure S6. Kaplan-Meier analyses of TNBC patients. Kaplan-Meier plots of RFS (a and c) or OS (b and d) comparing the TNBC breast cancer patients with high (black) and low (gray) expression of B3GALT5 in tumor (a and b) and adjacent non-tumor parts (c and d) were analyzed. Figure S7. Knockdown of B3GALT5 inhibits mammosphere formation. AS-B634 cells transfected with control siRNA (si-Ctrl) or B3GALT5 siRNA (si-B3GALT5-1 and -2) and the representative pictures of mammosphere were shown. Scale bar: 250 μm. Figure S8. Quantification of lung metastasis. Lung sections were stained with H&E. Counting of the pre-metastatic colonies were based on the cell numbers per colony. Representative pictures were shown. Scale bar: 100 μm.

附加文件1：图S1。202例乳腺癌患者的临床及病理特征。
图S2：B3GALT5表达水平与乳腺癌分期及分级的相关性。针对Ⅰ级、Ⅱ级、Ⅲ级（a、c组）或Ⅰ期、Ⅱ期、Ⅲ~Ⅳ期（b、d组）患者的肿瘤组织（a、b组）及癌旁非肿瘤组织（c、d组），采用实时定量逆转录聚合酶链反应（quantitative real-time reverse transcription polymerase chain reaction, qRT-PCR）检测B3GALT5 mRNA的表达水平，统计分析采用单因素方差分析（one-way ANOVA）。
图S3：乳腺癌组织肿瘤部位的B3GALT5表达水平。采用GSE1456数据集绘制生存曲线，对比B3GALT5高表达（黑色）与低表达（灰色）患者的生存情况。
图S4：乳腺癌组织中B3GALT5高表达与不良临床结局（无复发生存期，relapse-free survival, RFS）相关。针对202例乳腺癌患者，分别分析其肿瘤组织（a~c组）及癌旁非肿瘤组织（d~f组）中B3GALT5高表达（黑色）与低表达（灰色）患者的无复发生存曲线：其中a、d组为Ⅰ~Ⅱ期患者，b、e组为Ⅰ~Ⅱ期且腔A型（Luminal A）乳腺癌患者，c、f组为Ⅰ~Ⅱ期且腔B型（Luminal B）乳腺癌患者。
图S5：乳腺癌组织中B3GALT5高表达与不良临床结局（总生存期，overall survival, OS）相关。针对202例乳腺癌患者，分别分析其肿瘤组织（a~c组）及癌旁非肿瘤组织（d~f组）中B3GALT5高表达（黑色）与低表达（灰色）患者的总生存曲线：其中a、d组为Ⅰ~Ⅱ期患者，b、e组为Ⅰ~Ⅱ期且腔A型乳腺癌患者，c、f组为Ⅰ~Ⅱ期且腔B型乳腺癌患者。
图S6：三阴性乳腺癌（triple-negative breast cancer, TNBC）患者的Kaplan-Meier分析。针对三阴性乳腺癌患者，分别分析其肿瘤组织（a、b组）及癌旁非肿瘤组织（c、d组）中B3GALT5高表达（黑色）与低表达（灰色）患者的无复发生存（a、c组）及总生存（b、d组）曲线。
图S7：沉默B3GALT5可抑制乳腺球（mammosphere）形成。将AS-B634细胞转染对照小干扰RNA（small interfering RNA, si-Ctrl）或B3GALT5小干扰RNA（si-B3GALT5-1、si-B3GALT5-2），并展示乳腺球形成的代表性图像。标尺：250 μm。
图S8：肺转移定量分析。采用苏木精-伊红染色（hematoxylin-eosin staining, H&E）对肺组织切片进行染色，根据每个转移集落的细胞数量计数预转移集落，并展示代表性图像。标尺：100 μm。