Fig 1: Theexpressionof Raftlinin nasal tissues. (A) The Raftlin levels from whole nasal tissue pieces in the control (n = 20), CRSwNP (n = 30), and CRSwNP + SK (n = 16) groups were analyzed byELISA assay. (B) Representative photomicrograph of nasal epithelium staining of Raftlin expression and its corresponding normal rabbit immunoglobulin G (IgG) as the isotype control for three different study groups. Yellow arrows: basal cells staining and red arrows: columnar cells staining. Scale bar = 50 μm. The quantitative intensity of staining for (C) the basal cells and (D) columnar cells was measured by Image-Pro Plus 6.0. * p < 0.05 vs. control; ** p < 0.01 vs. control. SK: smoking.
Fig 2: Identification of two core genes for LUAD. a Venn diagram showing two core genes by integration of gene expression, methylation and CNV data for LUAD. The differences in CNV (b), gene expression (c) and methylation (d) of CNTN4 between iC2 subtype and in iC1 subtype. (E) Kaplan-Meier survival analysis between high and low expression of CNTN4. The differences in CNV (f), gene expression (g) and methylation (h) of RFTN1 between iC2 subtype and in iC1 subtype. i Kaplan-Meier survival analysis between high and low expression of RFTN1
Fig 3: The correlation between IL-17 or TNF-α and the three measurable Raftlin levels (66 samples in total) was analyzed by Pearson R test. (A) Correlation between levels of IL-17 and Raftlin from whole nasal tissues, (B) between IL-17 and Raftlin from IHC intensity of basal cells, (C) between IL-17 and Raftlin from IHC intensity of columnar cells, (D) between levels of TNF-α and Raftlin from whole nasal tissues, (E) between TNF-α and Raftlin from IHC intensity of basal cells, and (F) between TNF-α and Raftlin from IHC intensity of columnar cells in all study subjects. IL: Interleukin; TNF-α: Tumor necrosis factor alpha; p < 0.05 was considered a statistically significant difference.
Fig 4: Overexpression of CNTN4 and RFTN1 suppresses migration and invasion of LUAD cells. a-d The migrated ability of LUAD cells transfected with CNTN4 and RFTN1 overexpression was assessed via wound healing assay. Magnification: 200×. e-h Transwell assay for the invasion of LUAD cells transfected with overexpression of CNTN4 and RFTN1. Magnification: 200×. **p < 0.01; ****p < 0.0001
Fig 5: CNTN4 and RFTN1 are down-regulated in LUAD and their overexpression inhibits proliferation of LUAD cells. a-c Western blot for the expression of CNTN4 and RFTN1 proteins in LUAD and normal tissue specimens. Western blot confirmed that (d, e) CNTN4 and (f, g) RFTN1 were successfully overexpressed in LUAD cells. (Full-length blots/gels are presented in Supplementary Fig. 1). h-j Clone formation assay for the proliferation of LUAD cells transfected with overexpression of CNTN4 and RFTN1. Magnification: 200×. ****p < 0.0001
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