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. Author manuscript; available in PMC: 2017 Jan 1.
Published in final edited form as: Methods Enzymol. 2015 Sep 26;569:3–22. doi: 10.1016/bs.mie.2015.08.008

Figure 2. Proximity dependent biotinylation by BioID-LaA.

Figure 2

Control (non-transfected) human diploid foreskin fibroblast cells (BJ) or those cells stably expressing BioID-only or BioID-LaA (myc-BioID-LaA) by retroviral transduction were analyzed 18 hours after incubation in 50 μM biotin. (A) By IF analysis the BioID-only localizes to the nucleus and cytoplasm, whereas BioID-LaA (both red) localizes primarily to the nuclear envelope and minimally within the nucleoplasm. Biotinylated proteins, detected with labeled streptavidin (green), co-localize with the fusion proteins (red). DNA is labeled with Hoechst (blue). Bar, 10 μm. (B) IB analysis reveals migration of the fusion protein with anti-BioID and biotinylated endogenous proteins via streptavidin-conjugated HRP.