Fig. 1: CircDhx32 is upregulated in I/R-treated mouse hearts and H/R-treated cardiomyocytes.

a Heatmap showing numerous circRNAs with significant differential m⁶A modifications in I/R-treated mice. b, c RIP revealed the binding of circDhx32 with a m⁶A antibody in I/R-treated mice or H/R-induced cardiomyocytes. The Y-axis represents the percentage of input for each IP sample according to the formula: %Input =1/10*2^{Ct [IP] – Ct [input]}. n = 3. ^**P < 0.01 vs. Sham, ^*P < 0.05 vs.^. Ctrl. P values were determined via an unpaired t test. d Sanger sequencing was performed to validate that circDhx32 originated from its host gene. e Expression of linear RNA (Dhx32) and circDhx32 after Act D treatment for 24 h. n = 6. ^**P < 0.01 vs. Act D^-. P values were determined via an unpaired t test. f Relative RNA expression of GAPDH and circDhx32 in cardiomyocytes with or without RNase R incubation. n = 5. ^**P < 0.01 vs. Mock. P values were determined via an unpaired t test. g An RNA fractionation assay was used to evaluate the distribution of circDhx32 in the nucleus and cytoplasm under H/R conditions. n = 5. *P < 0.05 vs. Ctrl. P values were determined via an unpaired t test. h Subcellular localization of circDhx32 in H/R-induced cardiomyocytes was detected via FISH. The nuclei were stained with DAPI. Cy3 was used to stain for circDhx32. Scale bar = 20 µm. n = 4.