Extended Data Fig. 6: Validation of ERα as import shuttle for FKBP12.
From: Targeted protein relocalization via protein transport coupling
a. TRAM engaging ERα and FKBP12, raloxifene (10) unlinked control binds to ERα. b, FKBP12 nuclear import dose response derived from the analysis pipeline after a 3-hour treatment with 7 engaging ERα. c, Representative live-cell images of ERα-dependent FKBP12 import upon treatment with 7 or unlinked warheads 10 and 9 for 3 h. d, Nuclear localization of mCherry-FKBP12 after treatment with 7 or unlinked warheads 10 and 9 for 3 h. e, Nuclear localization of ERα after treatment with TRAM 7 or unlinked warheads 10 and 9 for 3 h. f, Nuclear mCherry localization upon treatment with 7 in competition with either 1x or 10x of 10 or 9 for 3 h. g, Nuclear localization of ERα upon treatment with 7 in competition with either 1x or 10x of 10 or 9 for 3 h. h, Representative live-cell images of the unlinked warhead competition assay for ERα-dependent FKBP12 import after a 3-hour treatment. Images in c, h are representative of three biological replicates. Data in b, d, e, f, g are mean ± s.d. of three independent experiments. Scale bars are 20 µm. P values in b, d, e, f, g, were determined by one-way ANOVA with Dunnett’s post hoc test comparing each condition to the DMSO control. Hoechst stains the nucleus.
