Fig. 4: CircACSL1 serves as a sponge of miR-8055 to regulate the expression of MAPK14.

A The expression level of MAPK14 was upregulated in MC patient samples and LPS-induced HCM inflammatory model as estimated by qRT-PCR and WB. B The expression level of miR-8055 was downregulated in MC patient samples and HCM inflammatory model as detected by qRT-PCR. C After transfection with sh-circACSL1 or circACSL1 overexpression vector, the miR-8055 expression was measured by qRT-PCR. D The mRNA level of MAPK14 in HCM with overexpression or knockdown of circACSL1. E The mRNA level of MAPK14 in HCM with miR-8055 overexpression or knockdown. F The protein level of MAPK14 in HCM with overexpression or knockdown of circACSL1. G The protein level of MAPK14 in HCM with miR-8055 overexpression or knockdown. H, I The higher expression of MAPK14 in HCM with circACSL1 overexpression could be rescued by miR-8055 overexpression, and sh-miR-8055 could relieve the inhibition of sh-circACSL1 on MAPK14 expression. J, K The expression of miR-8055 and MAPK14 could not be affected by mutant circACSL1. L1 miR-8055 was pulled down by the biotin-circACSL1 probe, detected by qRT-PCR. L2 circACSL1 was pulled down by the biotin-miR-8055 probe, detected by qRT-PCR. The relative level was normalized to input. M RNA-FISH revealed the cytosolic colocalization of circACSL1 and miR-8055 in HCM. CircACSL1 probes were labeled with CY3 (red). miR-8055 probes were labeled with FAM (green). Nuclei were stained with DAPI (blue). Scale bar, 50 µm. N1 The complementary binding sites in wild-type (WT) and mutant sites (Mut) 3′-UTR of MAPK14 and circACSL1 for miR-8055 targeting. N2 Luciferase reporter assay verified the molecular interactions of miR-8055 with the 3′-UTR of MAPK14 and circACSL1 wild-type transcripts. Data are presented as mean ±SD (n = 3 biologically independent samples). *p < 0.05; **p < 0.01; ***p < 0.001 (Student’s t test).