Nonlinear Bivariate Associations and Mononuclear Cell‐Type‐Specific Expression Level Differences in the STING Signalling Pathway

Figures

• 

  FIGURE 1. Representative histograms for each analyte and each cell type demonstrates definitive expression levels.

• 

  FIGURE 2. Cell‐type‐specific expression levels of STING pathway components. Expression was assessed by signal amplification and flow cytometry, and median expression levels, as indicated by fluorescence intensity, were converted to Z scores per analyte to facilitate comparisons. The prefix p in analyte names indicates the phosphorylated substrate. Error bars represent the standard error of the mean. Under each plot, the ANOVA analyses are shown for each pair of cell types. The p‐values shown have been adjusted by the Bonferroni correction for multiple comparisons.

• 

  FIGURE 3. Cell‐type‐specific regulation of the type I interferon branch of the STING pathway. Bivariate expression levels are shown. (a) STING pathway model showing the type I interferon and the NFκB branches. (b) Bivariate relationships of STING with BDNF and phospho‐Akt are plotted for the various mononuclear cell types. Linear regressions with corresponding r values are shown. (c) Bivariate relationships of phospho‐STING with BDNF and phospho‐Akt are plotted for B lymphocytes. Logarithmic regressions are shown with the corresponding equation in each upper panel. The lower panels plot the BDNF and phospho‐Akt with the natural log transformation of phospho‐STING with the linear regression and correlation coefficients shown in each panel.

• 

  FIGURE 4. Cell‐type‐specific bivariate relationships between phospho‐TBK1, phospho‐STING and phospho‐RelA. (a) The bivariate expressions of phospho‐TBK1 and phospho‐STING are shown with inverse power regression and the corresponding regression equation. (b) The bivariate expressions of phospho‐TBK1 and phospho‐RelA are shown with exponential regression and the corresponding regression equation. (c) The bivariate expression of the natural logarithmic transformation of phospho‐Rela and phospho‐TBK1 is shown with linear regression and the corresponding r values.

• 

  FIGURE 5. Non‐transformed versus ln transformed analyte correlation matrices for each mononuclear cell type are presented as a heat map. The bivariate relationships with r values > 0.6 and < −0.6 are shaded. Weak relationships are defined as correlations with 0.6 < r < 0.7 or − 0.6 > r > −0.7. Note that the correlation coefficient for ln(x) and y is not necessarily identical to the coefficient for x and ln(y). Also, the correlation coefficients for x versus ln(x) (signified by green fill in the figure) were > 0.9 with a single exception.

• 

  FIGURE 6. Cell‐type‐specific bivariate relationships between phospho‐TBK1 and phospho‐RelA. Additional aliquots of samples from 55 patients were assessed for the expression of phospho‐TBK1, phospho‐RelA and phospho‐IKKε in CD4+ T cells and monocytes. The bivariate expressions of phospho‐TBK1/phospho‐RelA and phospho‐TBK1/ln(phospho‐RelA) are shown with exponential trendlines. Exponential equations are shown in the top panels, and the correlation coefficients for the natural log‐transformed phospho‐RelA and phospho‐TBK1 relationships are shown in the middle panels.

• 

  FIGURE 7. STING agonist affects pathway components. Peripheral blood mononuclear cells from 6 healthy volunteers were treated with varying concentrations of the STING agonist diABZI (0 and 5 5‐fold dilutions from 1 μM) for one hour in culture medium at 37°C in an atmosphere of 5% CO2. (a) Phospho‐STING, phospho‐TBK1 and phospho‐RelA expression levels were analysed in (a) CD4+ T cells and (b) monocytes. Equations are shown for logarithmic and exponential models (upper panels). Log normal (Ln) transformations were performed for phospho‐STING and phospho‐RelA (lower panels), and the correlation coefficients (r) are shown.