Fig. 4. Circ_0006156 binds to and stabilizes S100A9 by blocking ubiquitination.
A The secondary mass spectrum result of S100A9 peptide. B, C Silver-staining assay after RNA pulldown showed that S100A9 binds to circ_0006156. D RIP assay showed that circ_0006156 binds to S100A9 in DU145 cells. E, F qRT–PCR and Western blot analyses showed that S100A9 protein, but not mRNA, was influenced after circ_0006156 knockdown or overexpression. G Western blot analysis of S100A9 expression after treatment with cycloheximide at the indicated time points in different groups. H S100A9 expression was increased after treatment with MG132. I Western blot analysis after co-IP showed that the ubiquitination level of S100A9 was decreased after overexpressing circ_0006156.