Journal: bioRxiv
Article Title: The synergistic effect of circRNA methylation promotes pulmonary fibrosis
doi: 10.1101/2021.08.05.455186
Figure Lengend Snippet: METTL3-mediated circRNA m 6 A modification promotes the activation of human pulmonary fibroblasts. A In situ hybridization detection of hsa_circ_0000672 and hsa_circ_0005654 colocalization with METTL3. Scale=20 μm. B RIP test of the interaction of METTL3 with hsa_circ_0000672 and hsa_circ_0005654. C Changes in circRNA m 6 A modification after METTL3 knockdown were detected by me-RIP. D PCR was used to detect the changes in hsa_circ_0000672 and hsa_circ_0005654 expression after knocking down METTL3. E Hsa_circ_0000672 and hsa_circ_0005654 with the m 6 A site mutated or wild-type overexpression plasmids were constructed. F-G The overexpression efficiency of wild-type and m 6 A site-mutated overexpression plasmids in human pulmonary fibroblasts was detected by PCR. H-K WB was conducted to detect the effect of the wild-type overexpression plasmid on the human pulmonary fibroblast activation markers FN1, Collagen 1 and α-SMA. I-O WB was conducted to detect the effect of the m 6 A site mutated overexpression plasmids on the human pulmonary fibroblast activation markers FN1, Collagen 1 and α-SMA. The data are presented as means±SD, *P < 0.05, ** P < 0.01, *** P < 0.001.
Article Snippet: First, an Arraystar m 6 A-circRNA Epitranscriptomic microarray was used to investigate the differences in m 6 A modification in circRNAs in the lung tissues of mice in the silicosis and control groups ( ).
Techniques: Modification, Activation Assay, In Situ Hybridization, Expressing, Over Expression, Construct, Plasmid Preparation