Journal: Frontiers in Immunology

Article Title: Altered circular RNA expressions in extracellular vesicles from bronchoalveolar lavage fluids in mice after bacterial infections

doi: 10.3389/fimmu.2024.1354676

Figure Lengend Snippet: Validation of individual circRNAs detected in microarray profiling using RT-qPCR. WT mice were exposed to Acid (0.1 N HCl) or LPS (1µg) for 1 day. BALF EVs and cells were collected and circRNAs were evaluated using qPCR (normalized to β-actin or GAPDH). (A) To validate the specific circRNAs using RT-qPCR, we first designed the specific primers targeting the BSJ region to detect circular forms of RNAs, as illustrated here (left panel). The specific sequence of each specific circRNAs is listed on the right panel. (B) Validation of specific circRNAs using RT-qPCR in BALF EVs. We selected five circRNAs that were highly altered in the BALF EVs using microarray profiling. Next, we validated the expression and alteration of each circRNA using RT-qPCR in BALF EVs. *p<0.05. (C) To confirm that the specific primers used to detect circRNAs fail to detect the linear form host RNA. Using circ30884 as an example, we confirmed that using the primer listed above, we failed to detect its host linear RNA XDH. The figures shown here represent repeats from three independent experiments. *p<0.05. (D) AMs and neutrophils were separately isolated from LPS-exposed mouse lung as the above. Circular RNAs were then detected using RT-qPCR. *p<0.05, ns, not significant.

Article Snippet: We first used the high-throughput RNA-sequencing microarray screening by Arraystar.

Techniques: Biomarker Discovery, Microarray, Quantitative RT-PCR, Sequencing, Expressing, Isolation