Additional file 1 of LINC00543 promotes colorectal cancer metastasis by driving EMT and inducing the M2 polarization of tumor associated macrophages
Creators: name: Zheng, Jinsen
name: Dou, Rongzhang
name: Zhang, Xinyao
name: Zhong, Bo
name: Fang, Chenggang
name: Xu, Qian
name: Di, Ziyang
name: Huang, Sihao
name: Lin, Zaihuan
name: Song, Jialin
name: Wang, Shuyi
name: Xiong, Bin
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Cell Biology
subject: Molecular Biology
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Information Systems not elsewhere classified
subject: Cancer

description: Additional file 1: Table S1. Relationship between LINC00543 and clinicopathological features of CRC patients(n=40). Table S2. The sequences of the primers for quantitative RT-qPCR. Table S3. The Sequences of siRNAs, Plasmid. Table S4. The Sequences of LINC00543 segmental antisense nucleotide fragment biotin probes. Fig. S1. LINC00543 is highly expressed in CRC tissues and is associated with poor prognosis of CRC patients. a. ISH analysis of the expression of LINC00543 in CRC tissues. LINC00543 is demonstrate as a long non-coding RNA without encoding proteins ability on chromosome 13 through online websites b. pubmed (https://pubmed.ncbi.nlm.nih.gov/), c. LnCAR (https://lncar.renlab.org/), and d. CPC2 (http://cpc2.gao-lab.org/). Fig. S2. LINC00543 promotes the invasion and metastasis of CRC cells. The construction of LINC00543 knockdown and overexpression stably transfected cell lines. a. The knockdown efficiencies of Si-LINC00543A, Si-LINC00543B and Si-LINC00543C in SW620 cells. b. The knockdown efficiencies of SH-LINC00543A and SH-LINC00543B in cells. c. The transfection efficiency of LINC00543 in SW480 cells. d. The Ki67 score of NC and SH cells. Fig. S3. LINC00543 regulates the EMT of CRC cells. The expression levels of E-cadherin, N-cadherin, and Vimentin in NC group and SH group nude mice. Fig. S4. LINC00543 regulates EMT by upregulating FOXQ1 in CRC cells. The expression levels of EMT-related transcription factors in SH and LINC00543 cells. Fig. S5. The Polysome analysis of SH cells. Fig. S6. LINC00543 indirectly regulates FOXQ1 by downregulating miR-506-3p. The interaction of LINC00543 with miR-506-3p and Pre-miR-506-3p were predicted through online prediction website. a. The interaction of LINC00543 with miR-506-3p was predicted through online prediction website STARBASE (https://starbase.sysu.edu.cn/). b. The interaction of LINC00543 with miR-506-3p was predicted through online prediction website LncBASE (https://diana.e-ce.uth.gr/lncbasev3/interactions). c. The LINC00543 and Pre-miR-506-3p interaction were predicted through bioinformatics tool miRanda. Fig. S7. LINC00543 inhibits the transportation of pre-miR-506-3p. a. The relative expression levels of miR-506-3p, Pre-miR-506-3p, and Pri-miR-506-3p were detected in LINC00543 cells. b. RAP were applied to analyze the interaction between LINC00543 and Pre-miR-506-3p. c. The expression of Pre-miR-506-3p in the nucleus and cytosol of LINC00543 cells. d. RIP analysis of XPO5 interaction with Pre-miR-506-3p in LINC00543 cells. e. RIP analysis of XPO5 interaction with LINC00543 in LINC00543 cells. f. Relative expression of XPO5 in SH and LINC00543 cells. g. Agarose gel electrophoresis analysis of regions of LINC00543 combination with Pre-miR-506-3p. h. RAP analysis of the interaction of four LINC00543 segments with Pre-miR-506-3p. i. RIP analysis of the interaction of Pre-miR-506-3p and LINC00543 segments in incubated of XPO5 cells. j and k. Analysis of the interactions of LINC00543, XPO5 and Pre-miR-506-3p in cells incubated with antisense nucleotide biotin probes of the 3 binding regions. Fig. S8. LINC00543 promotes macrophage recruitment and M2 polarization through regulating CCL2 expression. Transwell assays were used to verify the relationship between the expression of LINC00543 and macrophage recruitment. Fig. S9. LINC00543 promotes the tumorigenesis of CRC and the M2 polarization of macrophages in vivo. a. The expression of Ki67 in THP1+NC group and THP1+SH group. b. The expression of CD163 and foxq1 in THP1+NC group and THP1+SH group.