Extended Data Fig. 5: Signaling pathway analysis of murine organoids with KPT-330-mediated XPO1 inhibition.
a, Single-cell RNA-seq log normalized (transcripts per 10,000 – tp10k) expression of Xpo1 in control cells by cell type annotations. b, Violin plots of nuclear export signal (NES) containing gene module score in control cells by cell type annotations, scaled from 0 to 1. c, Single-cell RNA-seq log normalized expression of genes involved in MAPK, NFAT, AP-1, and Aurora kinase signaling in control cells by non-stem, and stem I / II / III annotations. d, LYZ secretion assay for organoids treated with KPT-330 (d) or KPT-8602 (e) for 6 days culture in ENR + CD media. Organoids were incubated in fresh basal media with 10 μM carbachol (Cch) for 3 h on day 6. Data normalized to ATP and standardized to the control in each experiment. Means and individual values are shown (N = 5), dotted line represents the control value (1). One sample two-tailed t-test compared to 1; significant p values for (d) ordered left to right, **p = 0.0011, **p = 0.0049, **p = 0.0053, **p = 0.0098, **p = 0.0077, *p = 0.0156, for (e) ordered left to right, *p = 0.0132, **p = 0.0071, *p = 0.0295, *p = 0.0124. f, Single-cell RNA-seq log normalized expression of genes regulated by XPO1 signaling between KPT-330-treated and control cells by cell-type annotations. Color scale is relative to control, purple-to-grey increasing relative-expression. LYZ secretion assay for organoids treated with p53 modulators, inhibitor Pifithrin-α (PFTa) and activator Serdemetan (Serd.) (g), CDK4/6 inhibitor Palbociclib (h), Aurora kinase inhibitor ZM447439 (i), over 6-day culture in ENR + CD media with or without 160 nM KPT-330. Organoids were incubated in fresh basal media with 10 μM carbachol (Cch) for 3 h on day 6. Data normalized to ATP and standardized to control in each experiment. Means and individual values are shown (N = 5), and the dotted line represents the control value (1). One sample two-sided t-test compared to 1; significant p-values for (i) ordered left to right, **p = 0.0073, *p = 0.0259, *p = 0.0151, **p = 0.0040, *p = 0.0331.
