Extended Data Fig. 3: NMNAT1 can be utilized as a nuclear import shuttle.
From: Targeted protein relocalization via protein transport coupling
a, Representative live-cell images of clonal import line A (IL-A) upon treatment with 1 or small molecule controls for 3 h. b, Quantification of localization upon treatment of the IL-A with unlinked warheads 2 and 3. c, Comparison of the relative mCherry/GFP median fluorescence intensity ratio between the three isolated clonal import lines (IL-A-C). d, Ability of three representative import cell lines (IL) with different relative target (mCherry) and shuttle (NMNAT1) ratios to redistribute a diffuse protein target after 3-hour treatment with 1. e, Representative live-cell images of clonal import line B (IL-B) upon treatment with 1 for 3 h. f, Representative live-cell images of clonal import line C (IL-C) upon treatment with 1 for 3 h. g, Quantification of NMNAT1 protein localization change upon treatment with 10 nM 1 in IL-A-C possessing different mCherry/GFP ratios. h, Levels of GFP after treatment with 1 for 3 h in four isolated import cell lines with varying nuclear protein expression levels. i, Levels of mCherry after treatment with 1 for 3 h across four isolated import cell lines with varying diffuse protein expression levels. Images in a, e, f, are representative of three biological replicates. Data in b, c, d, g, h, i are mean ± s.d. of three independent experiments. Scale bars are 20 µm. MFI = median fluorescence intensity. P values in h, i were determined by two-way ANOVA with Šidák post hoc test comparing the treated condition to the DMSO control for each line. P values in b, d were determined by unpaired two-tailed t-tests comparing treatment to the DMSO control.
