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Fig. 1 | Acta Neuropathologica Communications

Fig. 1

From: C9orf72-associated SMCR8 protein binds in the ubiquitin pathway and with proteins linked with neurological disease

Fig. 1

Protein interaction analyses by Western blotting and co-IP of the SMCR8 complex in HEK 293T cells (see Fig. S1 for antibody analyses). a Endogenous C9-L (arrow) co-IPs with FLAG-tagged SMCR8. The thick arrowhead marks a band consistent in size with C9-S. b FLAG-tagged C9orf72 co-IPs both endogenous and co-transfected HA-tagged SMCR8. c FLAG-tagged WDR41 protein co-IPs both endogenous C9orf72 and SMCR8 proteins (indicated by arrows). Tagged C9orf72 and WDR41 proteins of (b) and (c) are not visible in whole cell lysates at the Western blot film exposure times shown. d C9orf72-FL, FL-SMCR8, and empty vector were immunoprecipitated on α-FLAG agarose from transfected 293T whole cell lysates, resolved on a polyacrylamide gel, and silver-stained. IP reactions were in the presence or absence of 50 μg/ml RNases. Complex immunoprecipitate samples were analyzed by MS sequencing. Arrows indicate full-length protein bands. Protein molecular weight markers are those of Novex Sharp Pre-stained Protein Standard (Thermo Fisher Scientific)

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