Fig. 4: CircSPIRE1/ELAVL1/mRNA ternary complex stabilizes GALNT3 and QKI mRNA.

A Western blotting (left) as well as qPCR (right) validation of GALNT3 and QKI upon circSPIRE1 knockdown and overexpression. B Relative enrichment representing GALNT3 mRNA (left) and QKI mRNA (right) levels associated with circSPIRE1 junction compared to controls. Data represent mean ± S.D.; dot plots reflect data points from three independent experiments. The P values were determined by Student’s t test. C Reduced stability of GALNT3 (left) and QKI (right) upon circSPIRE1 knockdown. D Relative enrichment representing the enrichment of GALNT3 mRNA (left) and QKI mRNA (right) associated with the truncated ELAVL1 protein complex compared to an input control. IgG antibody served as a control. Data represent mean ± S.D.; dot plots reflect data points from three independent experiments. E IF-FISH assay showing that the colocalization of circSPIRE1/ELAVL1/mRNA was decreased upon knockdown of circSPIRE1. Scale bar, 10 µm. F Relative luciferase activity of luciferase reporter gene with GALNT3-WT or GALNT3-Mut (left) and QKI-WT or QKI-Mut (right). Data represent mean ± S.D.; dot plots reflect data points from three independent experiments. The P value was determined by Student’s t test. G Upon knockdown or overexpression of circSPIRE1, relative luciferase activity of luciferase reporter gene with GALNT3-WT or GALNT3-Mut (left) and QKI-WT or QKI-Mut (right). Data represent mean ± S.D.; dot plots reflect data points from three independent experiment. The P value was determined by Student’s t test. H Upon knockdown or overexpression of circSPIRE1, relative enrichment representing GALNT3 mRNA (left) and QKI mRNA (right) levels associated with ELAVL1 protein complex compared to an input control. IgG antibody served as a control. Data represent mean ± S.D.; dot plots reflect data points from three independent experiments.