Figure EV2. Biotinylated proteins evade purification from the AP step in APEAL.

Immunoblot analyses from lines expressing GFP‐TurboID‐HF or DCP1‐TurboID‐HF (denoted as GFP or DCP1, respectively) showing the presence of biotinylated proteins in the flow‐through after the AP‐step (for flow‐through 1, see DYNA‐IP here). The blot was overexposed to detect the faint streptavidin smear in flow‐through 1. The baits in these experiments undergo auto‐biotinylation, as reported previously. The results are representative of one experiment performed three times. Note that the immunodetectable biotinylation levels did not correlate well with the hits identified in Fig EV5 (compare GFP to DCP1). GFP and DCP1 are of similar molecular weight, while the bands below the upper band (~ 130 kDa) likely correspond to proteolytic cleavage products.