Fig. 6
The expression profiling of m6A circXpo6 and m6A circTmtc3 in pulmonary arterial smooth muscle cells (PASMCs) and pulmonary artery endothelial cells (PAECs) in hypoxia. a M6A levels of total circRNAs were determined based on colorimetric method in vitro. PASMCs and PAECs were exposed to 21% O2 and 1% O2 for 48 h, respectively. Total RNA was extracted and treated by RNase R. M6A levels were determined as a percentage of total circRNAs. b Schematic representation of exons of the Xpo6 and Tmtc3 circularization forming circXpo6 and circTmtc3 (black arrow). c RT-PCR validation of circXpo6 and circTmtc3 in PASMCs and PAECs exposed to 21% O2. Divergent primers amplified circRNAs in cDNA, but not in genomic DNA (gDNA). The size of the DNA marker is indicated on the left of the gel. d and e RT-PCR and qRT-PCR was performed after m6A RIP in PASMCs and PAECs exposed to 21% (N) and 1% O2 (H) for 48 h, respectively. Input was used as a control (d). IgG was used as a negative control (d and e). Values are presented as means ± SD. *P ≤ 0.05 (different from 21% O2 or the N-anti-m6A); **0.001 ≤ P ≤ 0.009 (different from the N-anti-m6A), (n = 3 each)