Fig. 4: BRRF2 is transferred from EBV-infected cells to macrophages via EVs and leads to the inhibition of innate immunity in recipient cells. | Nature Communications

Fig. 4: BRRF2 is transferred from EBV-infected cells to macrophages via EVs and leads to the inhibition of innate immunity in recipient cells.

From: Extracellular vesicles derived EBV tegument protein BRRF2 suppresses cGAS phase separation to promote anti-viral innate immune evasion

Fig. 4

A The box plot shows the differences in the immune and stromal cell compositions between the two groups of tumor samples with different BRRF2 expression levels based on the RNA-seq data. p values were determined via an unpaired two-side ttest. Center line, median (50th percentile); box bounds, 25 and 75th percentiles (interquartile range, IQR); whiskers, extend from the bounds to the smallest or largest value within 1.5 × IQR of the bound; minima and maxima, the end points of the whiskers. B Representative images of panCK (green), BRRF2 (yellow) and CD68 (orange) staining in NPC patients’ tumor tissues. Scale bars, 5 μm. C The average distance from BRRF2+ or BRRF2- tumor cells to CD68+ cells was measured from the IHC images. Each dot represents a patient’s tumor tissue. The experiment was performed as three independent biological replicates. **p < 0.01 according to a two-sided paired t test. D HPMs and THP1 cells were incubated with HEK293T-induced EVs carrying mCherry-BRRF2 for 4 h, and mCherry-BRRF2 was internalized into the cells. E HPMs and THP1 cells were incubated with HEK293T-induced EVs carrying mCherry-BRRF2 for 4 h. mCherry signals were detected by flow cytometry. The median fluorescence intensity (MFI) is shown. (Means ± SEMs of n = 3 experiments. ****p < 0.0001 according to a two-sided unpaired ttest). F EVs from the HEK293T-Flag-BRRF2 cell line inhibited cGAS-STING activation induced by HT-DNA in THP1 and HPM cells. THP1 cells and HPMs were incubated with EVs from the HEK293T-Flag-BRRF2 cell line for 2 h and then transfected with 2 μg of HT-DNA for 6 h. Cell lysates were collected for immunoblotting with the indicated antibodies. The experiment was performed as three independent biological replicates. Representative images was shown. Source data are provided as a Source Data file. G After an incubation with EVs and transfection with HT-DNA, total mRNAs were extracted from THP1 cells for a qPCR analysis of the expression of the IFNβ, RANTES and IL6 mRNAs. The error bars are derived from triplicate experiments. The data are plotted as the means ± SEMs. ns indicates not significant. ****p < 0.0001 according to two-way ANOVA followed by Sidak’s multiple comparisons test.

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