Skip to main content
. 2023 Nov 3;14:7048. doi: 10.1038/s41467-023-42774-6

Fig. 4. MMP21 promotes the non-cell autonomous basal invasion of APCMin/RasV12-transformed cells in vivo.

Fig. 4

a, b The non-cell autonomous upregulation of MMP21 in APCMin/RasV12 cells. a Immunofluorescence images of MMP21 in intestinal villi. APCMin-Villin-GFP, Villin-RasV12, or APCMin-Villin-RasV12 mice were injected with tamoxifen and were sacrificed 3 days later. Paraffin-embedded sections were stained with DAPI (blue), anti-GFP antibody (green), and anti-MMP21 antibody (red). b Immunofluorescence images of MMP21 in intestinal organoids from APCMin-Villin-GFP, Villin-RasV12, or APCMin-Villin-RasV12 mice at 24 h after tamoxifen treatment. Arrowheads indicate APCMin/RasV12-transformed cells with elevated MMP21 expression (a, b). c–h Effect of MMP21-knockout on basal extrusion of APCMin/RasV12 cells. c Strategy for the CRISPR-mediated MMP21-targeted mice. Physical maps of the murine MMP21 gene locus and sgRNA sequences are shown. The red line indicates the targeted region. Protospacer adjacent motif (PAM) sequences are underlined. d Genomic sequence of MMP21 locus from F1 generation MMP21-knockout mice. Note that 5890 bp covering Exon 1–Exon 7 of the MMP21 gene is deleted. e Western blot analysis of protein lysates prepared from intestinal tissues of MMP21(+/+), (+/−), (−/−) genotypes. β-actin served as a loading control. f Immunofluorescence images of MMP21 in intestinal villi from APCMin-Villin-RasV12 or APCMin-Villin-RasV12-MMP21 KO mice. Paraffin-embedded sections were stained with DAPI (blue), anti-GFP antibody (green), and anti-MMP21 antibody (red). Arrowheads indicate APCMin/RasV12-transformed cells. g, h Frequency of the basal extrusion of APCMin/RasV12 cells. g Immunofluorescence images of intestinal villi from APCMin-Villin-RasV12 or APCMin-Villin-RasV12-MMP21 KO mice 3 days after tamoxifen injection. An arrow indicates a basally extruded cell. Ba and Ap stand for the basal and apical sides, respectively (b, f, g). Scale bars, 50 μm (a) and 10 μm (b, f, g). h Quantification of apical and basal extrusion of transformed cells. Data are mean ± s.e.m. n = three independent experiments. *P = 0.0004, unpaired two-tailed t-test.