Fig 1: Down-regulation of SDC2 inhibits the FAK/ERK signaling pathway, and thus relieves atherosclerosis in mice. (A) Representative Western blots of SDC2 protein and its quantitation in the aortic lysate of mice after depletion of SDC2, normalized to GAPDH; (B) Representative Western blots of FAK, and ERK1/2 proteins and their quantitation in the aortic lysate of mice after depletion of SDC2, normalized to GAPDH; (C) Aortic plaque area determined using oil red O staining in mice after depletion of SDC2; (D) Lesion degree of aorta determined using HE staining in mice after depletion of SDC2 (400×); (E) The proliferation of collagen fibers measured using Masson’s trichrome staining in the aorta of mice after depletion of SDC2 (200×); and (F) Mac-3-labeled macrophages surrounding the aorta measured using immunohistochemistry in mice after depletion of SDC2 (400×); *p < 0.05 vs. mice treated with sh-NC; n = 8 for mice upon each treatment. All data were measurement data and expressed as mean ± standard deviation. Comparisons were analyzed using the unpaired t-test. The experiment was repeated 3 times.
Fig 2: Filtration of relevant pathways: (a) 52 pathways correlated with SDC2 were figured out using Pearson correlation analysis; 14 pathways were identified as bone metastasis associated using GSEA algorithm; and 10 were overlapped; (b) summarizations of GSEA results; (c) results of 10 pathways in Pearson correlation analysis; (d) results of specific pathways using GSEA algorithm.
Fig 3: SDC2 is highly-expressed in atherosclerosis. (A) A heatmap of the top 15 DEGs in ACS-related microarray data GSE19339. Abscissa indicates sample number and the ordinate indicates DEGs. Histogram in the upper right refers to color gradation and small squares in the figure represent the expression of a gene in one sample; (B) SDC2 expression patterns determined by RT-qPCR in the PBMCs isolated from patients with ACS (n = 86) and healthy individuals (n = 86), normalized to GAPDH; *p < 0.05 vs. healthy individuals; and (C) Representative Western blots of SDC2 protein and its quantitation in the aortic lysate of HFD-fed ApoE–/– mice (n = 40) and ND-fed ApoE–/– mice (n = 10), normalized to GAPDH; ND refers to ApoE–/– mice fed with normal diet and HFD refers to ApoE–/– mice fed with high-fat diet. *p < 0.05 vs. ND-fed ApoE–/– mice. All data were measurement data and expressed as mean ± standard deviation. Comparisons in panel (B) were analyzed using unpaired t-test, while comparisons in panel C were analyzed using paired t-test.
Fig 4: Identification of key transcription factors (TFs) in MESO: (a) expression of differently expressed TFs in patients with MESO; (b) volcano plot showed that 5 out of 318 TFs from the Cistrome database were differently expressed in MESO versus normal samples; (c) survival analysis of TCF7L1 (left) and SDC2 (right) in pancancer; (d) effect of expression levels of TCF7L1 and SDC2 on the survival status of patients with MESO.
Fig 5: Immune regulatory network and ATAC-seq validation. (a) Integrated network includingSDC2 and 4 TFs, 11 immune cells, and 10 pathways; (b) gene loci on different chromosomes; (c) intersection of different pick types (genic, intergenic, exon, upstream, intron, and distal intergenic); (d) distribution of binding loci relative to TSS; (e) correlation analysis of TCF7L1 and SDC2 (P < 0.001, R = 0.700); (f) in ATAC-seq data of MESO samples, multiple binding peaks were identified in SDC2 and TCF7L1 sequences.
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