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[Preprint]. 2023 Jul 11:2023.07.10.548456. [Version 1] doi: 10.1101/2023.07.10.548456

Fig. 2. NOTCH3+ mural cells underly meningeal tumorigenesis.

Fig. 2.

a, IHC for NOTCH3 in the adult human meninges showing expression is restricted to mural cells. Representative of n=3 biological replicates. Scale bars, 100μm and 10μm (insert). b, IF for NOTCH3 and the mural cell marker SMA in 3 adult human meningeal samples showing NOTCH3 is expressed in mural cells adjacent to smooth muscle cells in the meninges. DAPI marks DNA. Scale bar, 10μm. c, Experimental design for in vivo lineage tracing of NOTCH3+ mural cells during meningeal development (in utero recombination) or homeostasis (postnatal recombination). TAM, tamoxifen. d, Confocal microscopy of whole mount mouse convexity meningeal samples at P7, P30, or P90 after in utero recombination of the ROSAmT/mG allele showing NOTCH3 cells (green) are restricted to the perivascular niche during meningeal development. Representative of n=3 biological replicates per timepoint. DAPI marks DNA. Scale bar, 10μm. e, Confocal microscopy of whole mount mouse convexity meningeal samples at P30 or P90 after postnatal recombination of the ROSAmT/mG allele showing NOTCH3+ cells (green) are restricted to the perivascular niche during meningeal homeostasis. Representative of n=3 biological replicates per timepoint. DAPI marks DNA. Scale bar, 10μm. f, Experimental design for in vivo biallelic inactivation of Nf2 in NOTCH3+ cells during meningeal development (E16.5) or homeostasis (P30). Mice were monitored for 1 year after Nf2 inactivation. g, Coronal H&E images of 300μm decalcified mouse skull sections 1 year after treatment of mice with TAM. No gross tumors were identified, but insets show Nf2 inactivation in NOTCH3+ cells is associated with meningeal hyperproliferation after either in utero (E16.5) or postnatal (P30) treatment with TAM. Representative of n=5–8 biological replicates per condition. Scale bars 1mm.