(Novex), blotted onto an immobilon membrane using a Bolt Mini Blot Module (Novex), and probed with anti-Emi2 (this study), anti-cyclin B1 (Abcam; V152), anti-γ-tubulin (Sigma; T6557), anti-Tdrd3 (Cell Signaling Technology; D302G) and anti-Mad2 (Bethyl; A300-301A) antibodies. The crude extracts from mouse ovaries and rabbit reticulocyte lysates and the immunoprecipitates were separated by SDS-PAGE, blotted onto an immobilon membrane, and probed with anti-Pum1 (Bethyl; A300-201A), anti-HuR (SANTA CRUZ; sc-5261), anti-ELAVL2 (HuB) (Proteintech; 14008-1-AP), anti-FLAG (Sigma; F1804-1MG), anti-Tdrd3 (Cell Signaling Technology; D302G) and anti-DIG-AP (Roche) antibodies. The intensity of signals was quantified using ImageJ software.
+ Open protocol