Fig. 5: Slc1a3+ epithelial cells are not cancer-prone.
From: Pre-ciliated tubal epithelial cells are prone to initiation of high-grade serous ovarian carcinoma
a tdTomato+ tubal epithelial cells in mice containing Slc1a3-CreERT with floxed Trp53 and Rb1 genes, and an Ai9 reporter 1 and 360 days post induction (DPI) with tamoxifen. Hematoxylin and Eosin (HE) staining (left column) and immunostaining for tdTomato (arrows, brown color), Elite ABC method, hematoxylin counterstaining (right column). Scale bar, all images 200 μm. b PCR analysis of Trp53 and Rb1 gene structure in the same samples of microdissected cells from the tubal epithelium (TE, lanes 5–9) and lung neoplasm (LT, lane 4) of Slc1a3-CreERT Trp53loxP/loxP Rb1loxP/loxP Ai9 mice collected 1 year after tamoxifen induction. Samples with known gene structure (wild-type, WT, lane 1, floxed gene, L, lane 2, and recombinant gene, R, lane 3). 316-, 198-, and 163-bp fragments are diagnostic for floxed, excised, and wild-type alleles of the Trp53 gene, respectively. 295-, 269, and 247-bp fragments are diagnostic for floxed, excised, and wild-type alleles of the Rb1 gene, respectively. B, blank control (lane 10), M, DNA marker (Lane 11). Representative of 5 microdissection-PCR experiments. c Apoptotic cells (arrows) in the tubal epithelium 1 day after tamoxifen induction of Cre-mediated inactivation of Trp53 and Rb1 in Slc1a3-CreERT Trp53loxP/loxP Rb1loxP/loxP Ai9 mice (MUT TAM) and littermates without Slc1a3-CreERT (WT TAM). Dot-line rectangle indicates respective location of cells shown in the inset in the top image. TUNEL assay, methyl green counterstaining. Scale bar, 50 μm and 21 µm, inset. d Quantification of apoptotic cells 1 day after administration of tamoxifen (TAM+) or vehicle (TAM-) in Slc1a3-CreERT Trp53loxP/loxP Rb1loxP/loxP Ai9 mice (DM), Slc1a3-CreERT Trp53loxP/loxP Ai9 mice (p53) or littermates without Slc1a3-CreERT (WT). d One way ANOVA with the Tukey’s multiple comparison post hoc test (DM TAM+ vs. DM TAM-) *P = 0.0192, (DM TAM+ vs. WT TAM+) *P = 0.0123, (p53 TAM+ vs. DM TAM-) **P = 0.0035, (p53 TAM+ vs. WT TAM+) **P = 0.0022. Data are presented as mean values ± SD. Biological replicates (mice, n): DM TAM+ n = 3, p53M TAM+ n = 5, DM TAM- n = 4, WT TAM+ n = 4. Source data are provided as a Source Data file.
