Fig. 4. Clu binds the ribosome at the mitochondrial outer membrane.

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Fig. 4. Clu binds the ribosome at the mitochondrial outer membrane. Clu binds the ribosome at the mitochondrial outer membrane. (A) Western blot of transfected S2R+ cells showing Clu reciprocally co-immunoprecipitates with myc-tagged large (RpL7a) and small (RpS4, RpS7) subunit ribosomal proteins as well as eIF3g. Myc-tagged plant glucuronidase (GUS) was transfected and used as a negative control. (B) Western blot of fly extract from adults expressing GFP-traps for RpL30, RpL10Ab and eIF3-S9 showing Clu can also form a complex with these three proteins. Pull-down from the GFP-trap for the ribonucleoprotein Trailerhitch (Tral) was used as a negative control. Arrowheads designate the correct size proteins. The other bands are non-specific. (C) Approximate molecular weights of each GFP trap. (D) Western blot showing whole cell extract (WCE), cytoplasmic (cyto) and mitochondrial (mito) extract fractions from adult flies expressing UAS-RpL7a-myc. RpL7a is present in the 4× enriched mitochondrial fraction (mito-en). Calr, Calreticulin. (E) After performing reciprocal co-immunoprecipitations on both cytoplasmic and mitochondrial fractions of the same extract, Clu is present in the mitochondrial fraction, but not in the cytoplasmic fraction. y w extract is used as a negative control. The upper bands in the anti-myc blot are nonspecific. Aditya Sen, and Rachel T. Cox Biology Open 2016;5:195-203 © 2016. Published by The Company of Biologists Ltd