Fig. 2: CircFNDC3B impairs CD8+ T cell infiltration via CXCL10/11 suppression. | Communications Biology

Fig. 2: CircFNDC3B impairs CD8+ T cell infiltration via CXCL10/11 suppression.

From: Tumor cell-intrinsic circular RNA circFNDC3B attenuates CD8+ T cells infiltration in non-small cell lung cancer

Fig. 2

A Tumor volume of subcutaneous xenograft in NOD-SCID mice. A549 cells stably transfected with either vector control or circFNDC3B were injected subcutaneously. Purified CD8+ T cells from healthy volunteers were intravenously injected into the mice. Data were displayed as mean ± SD. (Student’s t-test, n = 6, ***P < 0.001). B Volcano plots depicting differentially expressed genes in A549 and H226 cells transfected with either vector control or circFNDC3B overexpression plasmid. Venn diagrams demonstrate the number of overlapping downregulated (C) and upregulated genes (D) between A549 and H226 cells. E Lollipop chart indicating the relative expression of overlapping genes based on RNA-seq analysis. F The analysis of online single-cell RNA sequencing databases TISCH2 showed that CXCL10 and CXCL11 are primarily expressed in macrophages, followed by tumor cells in the TME of NSCLC. G The basal expression levels of CXCL10 and CXCL11 in the culture supernatants of A549, CD45− cells, CD45+ cells, and CD14+ macrophages. Collectively, a total of 1 × 106 CD45− cells, CD45+ cells, and CD14+ macrophages were sorted by flow cytometry and cultured for 48 h. The culture supernatants were then applied for the measurement of CXCL10 and CXCL11. (one-way ANOVA, n = 3, ** P < 0.01, *** P < 0.001, **** P < 0.0001). H The proportion of CD45+ cells, CD45− cells, and CD14+ cells were measured by flow cytometry (one-way ANOVA, n = 3, * P < 0.05, *** P < 0.001). I Relative mRNA expression levels of CXCL10 and CXCL11 in A549 cells following circFNDC3B overexpression or knockdown, determined by RT-qPCR. Data were displayed as mean ± SD. (Student’s t-test, n = 3, *** P < 0.001). J Secreted CXCL10 and CXCL11 levels were quantified by ELISA. Data were displayed with mean ± SD. (Student’s t-test, n = 3, ** P < 0.01, *** P < 0.001). K Chemotaxis of CD8+ T cells toward the supernatants of A549 cells with circFNDC3B overexpression or knockdown was measured using Transwell assays. Data were displayed as mean ± SD. (Student’s t-test, n = 3, ** P < 0.01).

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