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Fig. 2 | Acta Neuropathologica Communications

Fig. 2

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

Fig. 2

Confirmation of proteins in the SMCR8 complex. Selected proteins detected in the SMCR8 interactome by MS sequencing were tagged and coexpressed with SMCR8 in HEK 293T cells. Most were found to specifically co-IP on α-FLAG agarose with FL-SMCR8 but not empty vector. Approximately 1% of the input lysate (lanes 1, 2) and 30% of the immunoprecipitate (lanes 4–7) were loaded on gels. IP reactions were in the presence or absence of 50 μg/ml RNases. Also included is a panel representative of tagged FL-SMCR8 protein present in the input and IP fractions (detected by α-FLAG antibody) and showing that RNase treatment did not affect SMCR8 immunoprecipitation (lower right). Test proteins were detected by α-V5 antibody, except FL-UBR5, which was detected by α-FLAG antibody (bottom right). The molecular weight of each test protein, including its epitope tag, is shown in brackets. Protein molecular weight markers are those of Novex Sharp Pre-stained Protein Standard (Thermo Fisher)

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