Extended Data Fig. 2: Illustration of gating and segmentation pictures automatically generated from analysis pipeline applied to three different cell types.
From: Targeted protein relocalization via protein transport coupling
a, Imaging analysis pipeline applied on HeLa cells. Gating histograms generated with the set bounds indicated. Masks for nuclear and cytoplasm areas overlayed onto respective fluorophore images. Cytoplasm areas are circumferentially grown from nuclei masks retaining the relative size between the masks. b, Imaging analysis pipeline applied to HEK293T cells. Masks for nuclear and cytoplasm areas overlayed onto respective fluorophore images, with segmentation of highly confluent cells and gating out of dead cells (high nuclear stain over a small area) and debris. c, Representative live-cell images for NMNAT1 relocalization in EL-A treated with 1 or control molecules in cells seeded at high (0.04 × 106 cells) and low density (0.01 × 106 cells). d, Quantification of nuclear GFP and mCherry from c. Images in c are representative of three biological replicates. Data in d are mean ± s.d. of three independent experiments. Scale bars are 20 µm. P values in d were determined by one-way ANOVA with Dunnett’s post hoc test comparing each condition to the DMSO control. Hoechst stains the nucleus.
