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Fig. 7. Association of MVH with MILI and MIWI. (A) Co-immunoprecipitation of MILI and MIWI with MVH. The 293T cells were transfected with plasmids that expressed FLAG-tagged MILI or MIWI and either Myc-tagged MVH or mock plasmids. The lysates were immunoprecipitated with the anti-Myc or anti-FLAG antibodies and detected with the anti-FLAG or anti-Myc antibodies. (B) Co-immunoprecipitation of MILI and MIWI with MVH in the testis lysate. Testis lysate of normal mice was immunoprecipitated with pre-immune serum or anti-MVH antibody, and detected with the anti-MILI-26F antibody recognizing both MILI and MIWI. (C) Schematic structure of the truncated MVH protein. Broken boxes show the helicase domains. E, EcoRI; H, HindIII; Xh, XhoI. (D) CBB staining of the GST-fusion proteins. GST-fusion proteins that were purified with glutathione-Sepharose 4B were loaded on 4-20% SDS-PAGE. (E) GST pull-down assay. Testis lysates were incubated with GST or the various GST-MVH fusion proteins. The MILI and MIWI proteins that were pulled down with glutathione-Sepharose were detected by western blotting using antibody 26F.





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