Figure 3.

miR-9 and miR-181a regulate Bim ubiquitination-dependent degradation. A: Ubiquitination of Bim in RAW264.7 cells. Proteins immunoprecipitated with anti-Flag antibody were immunoblotted with an anti-HA antibody. Ubiquitinated Bim was detected as upper-shifted bands in anti-ubiquitin blotting. Ubiquitinated Bim was detected when cells were treated with MG132, and marked enhancement of its ubiquitination and reduction of total amount was induced by miR-9 and miR-181a transfection. B: qRT-PCR was used to detect the overexpression and downregulation of miR-9 and miR-181a. C: The amounts of soluble Cbl and Bim in the synovial fluid samples by using enzyme-linked immunosorbent assay.