https://orcid.org/0000-0003-4042-8290
Figures & data
Table 1. Top 100 genes co-expressed with F5 in tumors of the Scandinavian breast cancer cohort and the TCGA breast cancer cohort.
Figure 1. GO analysis of the F5 co-expressed genes in breast tumors.
Bar plots depicting the – log10 P-values for the most significant GO terms identified in the Scandinavian breast cancer cohort (A) and the TCGA breast cancer cohort (B). Functional annotation clustering analysis of the top 100 F5 co-expressed genes was performed using DAVIDCitation23 and REVIGO.Citation14 The representative terms (blue bars) are indicated with CAPITAL letters followed by the redundant, but more specific terms (gray bars).
Figure 2. T cell activation marker CD69 levels, WST-1 proliferation analysis and cell migration of FV treated Jurkat T cells.
Cell surface expression of CD69, measured by flow cytometry, following 24 hour treatment with human FV (7 µg/mL) or control (50% (vol/vol) glycerol/H2O) in Jurkat T cells. The T cell mitogen PHA (20 µg/mL) served as positive control for CD69 expression. Left: Relative cell surface expression of CD69. Mean values + SD (n = 6 biological parallels) of two individual experiments. Right: Representative histogram showing the cell surface expression of CD69 for isotype control (background staining), control, human FV, and PHA (positive control). B) Cell viability following 24, 48, and 72 hours of treatment with human FV (7 µg/mL) or control (50% (vol/vol) glycerol/H2O) in Jurkat T cells. WST-1 was added to the cells and OD measurements were conducted after 1 hour incubation. Mean values + SD (n = 15 biological parallels) of three individual experiments. C) The relative number of migrated Jurkat T cells in response to human FV (7 µg/mL) or control (50% (vol/vol) glycerol/H2O) in the bottom chamber of a transwell assay. CXCL12 served as a positive control for migration. Cells were allowed to migrate for 24 hours. Mean values + SD (n = 9 biological parallels) of three individual experiments. P-values for two-group comparisons (Mann-Whitney U) are indicated.
Figure 3. Correlations between F5 tumor expression and breast cancer specific gene modules.
Bar chart showing Spearman correlation between F5 tumor expression and the eight co-expressed gene modules representing breast cancer specific transcriptional programs.Citation24 P-values are indicated for significant associations. Data obtained from the 1881-sample data set in GOBO.Citation15
Figure 4. F5 tumor expression according to the degree of TILs.
Box and whisker plot showing the distribution of F5 mRNA expression in tumors with low and high degree of TILs. Data from the Scandinavian breast cancer cohort (n = 44) were used. P-value for two-group comparison (Mann-Whitney U) is indicated.
Figure 5. Tumor abundance of subsets of leukocytes according to F5 expression.
A) Average relative fractions of 22 leukocyte subtypes according to low (<3rd quartile) and high (>3rd quartile) tumor expression of F5 in the Scandinavian breast cancer cohort (n = 131) and TCGA (n = 981). Data from CIBERSORT analysis. B) Relative abundance of cells for the leukocyte subtypes showing significantly different abundance between low and high F5 tumor expression.
Table 2. Correlations between F5 and gene markers of immune cells in the TCGA Breast cancer population in TIMER18.
Figure 6. Correlation between F5 tumor expression and tumor purity in breast cancer.
Scatterplot showing Spearman’s correlation between F5 tumor expression and tumor purity in the TCGA Breast Invasive Carcinoma population. Analyzed by TIMER.Citation18 The gene expression level of F5 was displayed with log2 RSEM on the y-axis and tumor purity on the x-axis.
Figure 7. Relapse-free survival in breast cancer patients stratified by F5 expression in tumors.
Kaplan-Meier survival curves and upper quartile survival times for 10-year relapse free survival, stratified according to high (red curves) and low (black curves) F5 gene expression in all tumors and molecular subgroups. Data derived from the Kaplan–Meier plotter database.Citation17 Vertical tick‐marks show censored individuals. HR = hazard ratio.
Figure 8. Multivariate analysis of relapse-free survival in breast cancer patients stratified by F5 expression in tumors.
Left: Kaplan–Meier plots for 10‐year relapse-free survival in basal‐like breast tumors (n = 115) from the Gene expression based Outcome for Breast cancer Online (GOBO),Citation15 stratified according to high (red curves) and low (gray curves) F5 gene expression. Vertical tick‐marks show censored individuals. Right: Corresponding multivariable Cox regression analysis for 10‐year relapse-free survival in basal‐like breast tumors from GOBO. HR = hazard ratio. n.a = not available.
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