FIGURE 1.

TurboID-Bok construct characterization. (A) V5-TurboID-Bok fusion constructs, T-Bok^WT and T-Bok^L34G. (B) Immunoblot for biotin-labeled species (detected with streptavidin/anti-streptavidin) in lysates from Bok KO HeLa cells transfected as indicated to express T-Bok^WT or T-Bok^L34G, without or with 2 h media supplementation with 50 μM biotin. Immunoreactivity of T-Bok constructs was assessed with either anti-Bok or anti-V5 (middle and lowest panels, respectively). (C) Anti-IP₃R1/IP₃R3 IP (lanes 1–3) and lysates (either pre- or post-IP; lanes 4–9) from Bok KO HeLa cells transfected as indicated, probed in immunoblots for the proteins indicated; p97 serves as a loading control. Co-migrating IgG heavy chain seen in the Bok probe of IPs is indicated by the asterisk. A 53kDa background band seen in the Bok probe of Bok KO cell lysates (lane 4) is indicated by the plus sign. Because Bok^L34G is relatively unstable (Schulman et al., 2016), to obtain equal expression, the amount of cDNA transfected for T-Bok^L34G was double that for T-Bok^WT.