Fig. 4: EMC4 and EMC7 are in close proximity to Rab7 and bind to LE-associated Rab7. | Nature Communications

Fig. 4: EMC4 and EMC7 are in close proximity to Rab7 and bind to LE-associated Rab7.

From: Selective EMC subunits act as molecular tethers of intracellular organelles exploited during viral entry

Fig. 4

a HEK 293 T cells expressing either Myc-BioID2, Myc-BioID2-EMC4, or EMC7-BioID2-HA were treated with biotin, and lysed with a RIPA buffer (containing 1% SDS). The resulting extract was subjected to affinity purification with Streptavidin C1 beads and the bound material eluted by a biotin-containing elution buffer. The input and eluted materials were subjected to SDS-PAGE and immunoblotted with indicated antibodies or probed with Streptavidin-HRP (to detect the total biotinylated proteins in each sample). b HEK 293 T cells were lysed and the resulting extract was incubated with either the control IgG or EMC4 antibody. The precipitated material was subjected to SDS-PAGE followed by immunoblotting with the indicated antibodies. c HEK 293 T cells transfected with the indicated plasmid were lysed, the resulting extract was incubated with EMC4 antibody, and the precipitated material subjected to SDS-PAGE and immunoblotting with the indicated antibodies. d–e HEK 293 T cells transfected with the indicated siRNA and DNA construct were lysed and the resulting extract was incubated with either an EMC4 (d) or FLAG antibody-conjugated agarose beads (e). The precipitated material was subjected to SDS-PAGE and immunoblotted with the indicated antibodies. All experiments were independently repeated three times. Source data are provided as a Source Data file.

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