SEC6 Antibody (9H5)

Western Blot: SEC6 Antibody (9H5) [NBP1-97500] -

Western Blot: SEC6 Antibody (9H5) [NBP1-97500] - Establishment & validation of tagged exocyst subunit cell lines. a Schematic showing a region of subunit gene targeted by sgRNA & tags to be inserted C-terminally, in-frame with the coding region. b Strategy to isolate CRISPR/Cas9-mediated sfGFP-tagged clones of exocyst subunits in NMuMG cells. c Western blots with subunit specific antibodies show successful incorporation of sfGFP in both alleles (EXO70, SEC5, & SEC8), or in one allele (SEC3 & SEC6). A shRNA specific to SEC3 was used to confirm the identity of multiple bands in the blot. d Confocal images of live wild-type NMuMG cells & endogenously tagged SEC3-GFP, SEC5-GFP, SEC6-GFP, SEC8-GFP, & EXO70-GFP cell lines. Scale bar = 10 μm. e Protein–protein interaction heat map for endogenous SEC3-GFP, SEC5-GFP, & EXO70-GFP pulldown using GFP-Trap nanobodies & MS. Yellow indicates baits, blue the protein identified with high confidence, & gray denotes preys that were not identified. The experiments were repeated three times for EXO70-GFP & twice for SEC3-GFP & SEC5-GFP. All experiments were also partially confirmed by IP–WB in at least three independent experiments, with similar results. f Western blot analysis to assess the relative abundance of exocyst subunits fused to sfGFP, using anti-GFP antibodies (SEC3 & SEC6 quantifications corrected for heterozygosity). GAPDH was the loading control. Y axis shows the ratio of GFP/GAPDH. Quantification shows pooled data from three experiments Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30510181), licensed under a CC-BY license. Not internally tested by Novus Biologicals.