Journal: Cell Death Discovery

Article Title: WWP1 gain-of-function drives developmental anoikis through TGFβ pathway during neurodevelopment

doi: 10.1038/s41420-026-02977-4

Figure Lengend Snippet: A Scheme of WWP1 variant-expressing cell line establishment and downstream analysis. The image is created with Biorender.com. B Representative images of dead cell population in WWP1 variants-expressing HeLa cell lines treated with gefitinib. Red, propidium iodide (PI). Scale bar, 200 μm. C Quantification of the PI + counts in HeLa cell lines treated with gefitinib. n = 3 for each condition. Two-way ANOVA with Tukey’s post hoc test. * p < 0.05; ** p < 0.01. D Functional GO enrichment analysis of upregulated and downregulated DEGs between WWP1 E798V and control cell lines. The bar color indicates the cluster categories of the GO term. BP, biological process; CC, cellular component. E Differential PROGENy pathway activity scores of WWP1 E798V relative to control. The pathway with the lowest value, TGFβ, is indicated by a deep blue color. F Gene set enrichment plots for TGFβ signaling and epithelial-mesenchymal transition. NES, normalized enrichment score. G Western blot analysis of phospho-SMAD2 and SMAD2 in WWP1 variant-expressing HeLa cell lines treated with vehicle or hTGFβ1 (10 ng/mL). ACTIN is used as the loading control. H Quantification of normalized pSMAD2 to SMAD2 protein expression ratio in WWP1 variant-expressing HeLa cell lines. n = 5. Kruskal-Wallis test with Dunn’s post hoc test. * p < 0.05; ns not significant. Bar graphs indicate mean ± SEM.

Article Snippet: Non-cytotoxic dosing conditions were determined by the cell viability using Propidium Iodide/RNase staining solution (Cell Signaling Technology, #4087).

Techniques: Variant Assay, Expressing, Functional Assay, Control, Activity Assay, Western Blot