Fig. 6: CircFndc3b promotes tumor progression by inhibiting CD8+ T cell infiltration in vivo. | Communications Biology

Fig. 6: CircFndc3b promotes tumor progression by inhibiting CD8+ T cell infiltration in vivo.

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

Fig. 6

A RNA FISH for circFndc3b in LLC cells. The positive controls for nucleus and cytoplasm were represented with U6 and 18S, respectively. CircFndc3b, U6, and 18S probes were labeled by Cy3. Nuclei were stained with DAPI. B The RNA-protein pull-down experiments with biotin-labeled sense or antisense circFndc3b probes in LLC cell lysates. The loading control was provided with GAPDH. C RIP assays with anti-TFII-I and anti-IgG antibodies were performed in LLC cell lysates. The precipitates were subjected to Western blot assay. The relative levels of TFII-I enriched and IgG-enriched circFndc3b were determined by RT-qPCR. Data were displayed as mean ± SD. (Student’s t-test, n = 3, **P < 0.01, ***P < 0.001). D Co-IP experiments using anti-TFII-I and IgG antibodies detected the interaction between TFII-I and STAT1 in LLC cells. E The relative mRNA levels of CXCL10 and CXCL11 in LLC cells overexpressing circFndc3b were determined by RT-qPCR. Data were displayed as mean ± SD. (Student’s t-test, n = 3, **P < 0.01, ***P < 0.001). F The tumor volumes were measured following subcutaneous injection of LLC cells with stable circFndc3b overexpression, knockdown, or vector control into C57BL/6 mice. Data were displayed as mean ± SD. (one-way ANOVA, n = 5, ** P < 0.01, *** P < 0.001). G The frequency of CD3+ CD8+ T cells in LLC tumors with either circFndc3b overexpression or knockdown was analyzed using flow cytometry. Data were displayed as mean ± SD. (one-way ANOVA, n = 5, *** P < 0.001). H IHC staining for CD8+ T cells in sections from LLC tumors with either circFndc3b overexpression or knockdown. I The proportion of IFN-γ+ CD8+ T cells was analyzed using flow cytometry. (one-way ANOVA, n = 5, ns: no significance). J The growth curves of the subcutaneous LLC tumor in C57BL/6 mice. Isotype or anti-PD-1 was administered intraperitoneally at the indicated time (arrows, Day 6, Day 10, Day 14, and Day 18). Data were displayed as mean ± SD. (Student’s t-test, n = 6, ***P < 0.001).

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