Additional file 1 of Wnt5a-induced M2 polarization of tumor-associated macrophages via IL-10 promotes colorectal cancer progression
Creators: name: Liu, Qing
name: Yang, Chaogang
name: Wang, Shuyi
name: Shi, Dongdong
name: Wei, Chen
name: Song, Jialin
name: Lin, Xiaobin
name: Dou, Rongzhang
name: Bai, Jian
name: Xiang, Zhenxian
name: Huang, Sihao
name: Liu, Keshu
name: Xiong, Bin
Publisher: name: figshare
Year: 2020
Subjects: subject: Biochemistry
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Science Policy
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Cancer

description: Additional file 1: Table S1. Correlation between the density of macrophages and clinicopathologic parameters (n = 63). Table S2. The sequences of the primers for RT-qPCR. Figure S1. Wnt5a+ TAM is significantly associated with prognosis in CRC patients. (a) Representative IHC staining of Wnt5a in CRC sample. Bar = 100 μm. (b) Wnt5a+ TAMs expression was significantly elevated in primary CRC tissues compared with normal colorectal tissues. Error bars, SEM. Statistical analysis was conducted using one-way ANOVA. (c) Correlation analysis between Wnt5a+ TAMs expression and RFS of CRC patients. (d) Correlation analysis between Wnt5a+ TAMs expression and OS of CRC patients. (e) Association of TAMs expression with RFS of CRC patients. (f) Association of TAMs expression with OS of CRC patients. ***P &lt; 0.001. Figure S2. (a) Relative expression of Wnt5a mRNA in M0, M1, M2 macrophages and TAMs cocultured with HCT116 or DLD-1. Error bars, SEM. (b) Western blot analysis of Wnt5a expression in CRC cell lines and CRC cell lines co-cultured with macrophages. (c) Western blot analysis of the level of p-CaKMII in Wnt5a-treated M0 macrophages. All experiments were performed in triplicate. Statistical analysis was conducted using one-way ANOVA. ns, not significant. ***P &lt; 0.001. Figure S3. Wnt5a does not directly influence the malignant biological behavior of CRC cells. (a) RT-qPCR analysis of Wnt5a expression in SW480 cells transfected with sh-Wnt5a or sh-NC. (b) CCK-8 assay analysis of cell viability in SW480 cells transfected with sh-Wnt5a or sh-NC. (c) Representative photographs and quantification of clone formation assay in SW480 cells transfected with sh-Wnt5a or sh-NC. Bar = 0.5 cm. (d) Transwell migration assay analysis of SW480 cells transfected with sh-Wnt5a or sh-NC. Bar = 200 μm. (e, f) CCK-8 assay analysis of cell viability in HCT116 or DLD-1 cells treated with Wnt5a. (g, h) Representative photographs and quantification of clone formation assay in HCT116 or DLD-1 cells treated with Wnt5a. Bar = 0.5 cm. (i, j) Transwell migration assay analysis of HCT116 or DLD-1 cells treated with Wnt5a. Bar = 200 μm. Error bars, SEM. All experiments were performed in triplicate. Statistical analysis was conducted using Student’s t test. ns, not significant. *P &lt; 0.05. ***P &lt; 0.001. Figure S4. (a) RT-qPCR analysis of Wnt5a expression in M0 macrophages, TAMs and TAMs transfected with sh-Wnt5a or sh-NC. Error bars, SEM. (b) Western blot analysis of Wnt5a expression in M0 macrophages, TAMs and TAMs transfected with sh-Wnt5a or sh-NC. All experiments were performed in triplicate. Statistical analysis was conducted using one-way ANOVA. ns, not significant. ***P &lt; 0.001.