Fig. 1: Identification and distribution of circFNDC3B. | Communications Biology

Fig. 1: Identification and distribution of circFNDC3B.

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

Fig. 1

A The outside circle represents gDNA, while the inside circle displays the back-spliced junction reads of circRNAs for each sample. Different sample was represented by different color. B The number of back-spliced reads and detected circRNAs was quantified in four paired normal and tumor tissues of NSCLC patients. C A heatmap illustrates the differentially expressed circRNAs between the four paired normal and tumor tissues in NSCLC. The columns represent individual tissue samples, while rows correspond to distinct circRNAs. D The expression levels of circFNDC3B were measured in 100 paired normal and tumor tissues in NSCLC. Data are displayed as mean ± SD. (Student’s t-test, ***P < 0.001). E NSCLC patients were grouped into circFNDC3B low expression cohort (n = 50) and high expression cohort (n = 50) based on the median expression level. Kaplan-Meier survival curves were used to compare overall survival between the two groups (Log-rank test). F Sequencing analysis confirmed the head-to-tail back-splicing junction of circFNDC3B. G Convergent and divergent primers were used to amplify circFNDC3B and GAPDH from cDNA and gDNA in A549 cells. H The relative RNA levels of GAPDH, circFNDC3B, and linear FNDC3B (mFNDC3B) were assessed in A549 and H226 cells, treated with or without RNase R. Data were displayed as mean ± SD. (Student’s t-test, n = 3, ***P < 0.001, ****P < 0.0001). I The stability of circFNDC3B and mFNDC3B was assessed in A549 cells at different time points following Actinomycin D treatment (n = 3). J RT-qPCR analysis was performed to determine the subcellular distribution of circFNDC3B in the nucleus and cytoplasm of A549 and H226 cells. U1 and GAPDH served as nucleus and cytoplasmic marker, respectively (Student’s t-test, n = 3, **P < 0.01, ****P < 0.0001). K RNA FISH for circFNDC3B in A549 cells. The positive controls for nucleus and cytoplasm were represented with U6 and 18S, respectively. CircFNDC3B, U6, and 18S probes were labeled with Cy3. Nuclei were stained with DAPI.

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