Additional file 6 of Exosomal miR-106a-5p from highly metastatic colorectal cancer cells drives liver metastasis by inducing macrophage M2 polarization in the tumor microenvironment
Creators: name: Liang, Yahang
name: Li, Junyu
name: Yuan, Yuli
name: Ju, Houqiong
name: Liao, Hualin
name: Li, Mingming
name: Liu, Yang
name: Yao, Yao
name: Yang, Lingling
name: Li, Taiyuan
name: Lei, Xiong
Publisher: name: figshare
Year: 2024
Subjects: subject: Biophysics
subject: Biochemistry
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Cancer

description: Supplementary Material 6: Fig. S1 Single-cell expression atlas of CRC with or without liver metastasis. (A) Uniform manifold approximation and projection (UMAP) plot of the major cell types in all samples (n = 13). (B) Dot plot exhibiting the marker genes across the cell types. (C) Histogram showed the proportions of cell types in CRC tissues with or without liver metastasis. (D) UMAP plot of myeloid clusters. (E) Histogram showed the proportions of myeloid types in CRC tissues with or without liver metastasis. (F) Heatmap exhibiting the marker genes across macrophage subsets. (G) Histogram showed the proportions of macrophage types in CRC tissues with or without liver metastasis, Fig. S2 Transwell assay was conducted to explore the metastatic capability of four different CRC cells. ** p &lt; 0.01, *** p &lt; 0.001, Fig. S3 (A) Western blot was employed to assess hnRNPA1 expression in samples obtained from miRNA pulldowns, utilizing nuclear, cytoplasmic, or exosomal lysates from SW620 cells. (B) RIP assays were conducted using an anti-hnRNPA1 antibody (or IgG as a control) on lysates derived from SW620 cells or exosomes. qRT-PCR was employed to quantify miR-106a-5p levels in the immunoprecipitated samples, expressed as percentages relative to the input (% input). *** p &lt; 0.001, Fig. S4 (A) qRT-PCR was performed to confirm the efficiency of lentivirus-mediated knockdown of miR-106a-5p in HCT 116 cells. (B) qRT-PCR was performed to detect the expression of miR-106a-5p in the exosomes of HCT 116 cells treated with miR-106a-5p knockdown lentivirus and lentiviral control. (C) qRT-PCR was performed to confirm the efficiency of lentivirus-mediated overexpression of miR-106a-5p in SW480 cells. (D) qRT-PCR was performed to detect the expression of miR-106a-5p in the exosomes of SW480 cells treated with miR-106a-5p overexpression lentivirus and lentiviral control. E-F qRT-PCR was performed to detect the expression of miR-106a-5p in Mφ cells after transfection with miR-106a-5p mimics (E) and in M2 macrophages after transfection with miR-106a-5p inhibitor (F). ** p &lt; 0.01, *** p &lt; 0.001, Fig. S5 A-B Western blot was performed to detect the expression of SOCS6 in M2 macrophages after transfection with sh-SOCS6 (A) and in Mφ cells after transfection with SOCS6-OE (B). C-D Transwell assays were performed to assess the migration and invasion capabilities of HCT-8 (C) and LoVo (D) cells treated with various CM