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. 2024 Oct 29;221(11):e20231832. doi: 10.1084/jem.20231832

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© 2024 Hsu et al.

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Figure 3. — IMPA1/inositol maintains ALDH^high organoids with AR low expression from TRAMP mouse model. (A) The flow chart shows an experimental outline to demonstrate ALDH^high tumor organoids sorted from TRAMP mouse. (B) Dot plot of sorted ALDH^high and ALDH^low/− cell population from TRAMP mouse. (C) The phase contrast shows the images of sorted ALDH^high and ALDH^low/− organoids from TRAMP mouse. Scale bar, 50 μm. (D) Immunoblotting of ALDH^high and ALDH^low/− organoids from TRAMP mouse with indicated antibodies. Immunoblotting data were verified in at least two independent experiments. (E) Inositol levels were determined by K-INOSL assay kit. The inositol levels were normalized by protein concentration in each experimental group. Three independent experiments were performed. *, P < 0.05; **, P < 0.01 by two-tailed unpaired t test. (F) The phase contrast shows the images of ALDH^high organoids upon shLuc, shIMPA1 (#1 and #2). Scale bar, 50 μm. H&E staining and IHC analysis of ALDH^high organoids upon shLuc, shIMPA1 (#1 and #2) with indicated antibodies. Scale bar, 20 μm. (G) Quantification of ALDH^high organoids upon shLuc, shIMPA1 #1 and shIMPA1 #2. The data were shown as the mean ± SEM of three independent experiments for each group. **, P < 0.01 by two-tailed unpaired t test. Source data are available for this figure: SourceData F3.