![Ablation of Myct1 upregulates pathways involved in protein folding, ribonucleoprotein assembly, and mTORC1 signaling. (A) Experimental workflow of the scRNA-seq experiment. CD45negCD31+ cells were sorted from mesenteric fat of 12-wk-old wild-type and Myct1ecKO mice. n = 2 per genotype. (B) UMAP representation of cells retained in the scRNA-seq and colored according to cell type. BECs, blood endothelial cells; LECs, lymphatic endothelial cells; vSMCs, vascular smooth muscle cells. (C)Myct1 is efficiently deleted from ECs in Myct1ecKO mice. Distribution of Myct1, Pecam1, and Cdh5 expression (ln[normalized counts +1]) in the BEC cluster in wild-type and Myct1ecKO cells. (D) GO terms and Hallmark gene sets overrepresented among upregulated (red bars) or downregulated (gray bars) genes in Myct1ecKO BECs compared with wild-type BECs. (E)Myct1 deficiency in ECs increases protein levels of mTORC1 target Mat2a (methionine adenosyltransferase 2A). Staining of WAT of Myct1ecKO or wild-type control mice for Mat2a (green) and pan-endothelial marker Erg (magenta). Scale bar, 50 µm. (F) Quantification of data shown in E. n = 8 mice per genotype from three independent experiments. mean ± SD; paired t test, P = 0.0191 (*). Icons used in A were created with BioRender.com and modified in Affinity. See also Fig. S3, A–D. Refer to the image caption for details.](https://cdn.rupress.org/rup/content_public/journal/jem/223/5/10.1084_jem.20251497/2/jem_20251497_fig3.png?Expires=1779627154&Signature=y0b5Bmdf7IX9xEhijK1aEzwvAiERbOKunJUfRfMmiEwc5CleFoq659B0XjnuJH8CdrZrObKK0QLjkKDuilFEEq4OMl2sV0AelRkR-M2wXVgfDeeriBQtAkDnY46mZoJA80~vCZw9j-2M6zsoXyFFHVkhXd0rOta-7d-mZjfCxU9Ge9scGN-rVzvL7XCQm0x~nMcr~p-lqnvlD8t9hQZ~4BfpI70oCXAH9quPJ4bl0gU9SJ36AW37lK9ZH~hsm2wuCDZ2hvVzfqZTmLZILA6onpBC-Mumed11n4auXqY-kAK9~ewKVjJPIzKykpX1nSfoglFFDdA12bpV5pALBVMCDA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Panel A shows a flowchart of the experimental workflow for the s c R N A-seq experiment, detailing the sorting of C D 45 n e g C D 31 plus endothelial cells from mesenteric fat of wildtype and M y c t 1 e c K O mice. Panel B presents a UMAP representation of cells colored according to cell type, including blood endothelial cells, lymphatic endothelial cells, pericytes slash vascular smooth muscle cells, mesothelial slash fibroblasts, and immune cells. Panel C displays violin plots showing the distribution of M y c t 1, P e c a m 1, and C d h 5 expression levels in the B E C cluster for wildtype and M y c t 1 e c K O cells. Panel D features bar graphs of Gene Ontology terms and Hallmark gene sets, highlighting pathways enriched among genes upregulated or downregulated in M y c t 1 e c K O B E C s. It shows enrichment of processes such as protein folding, regulation of the D N A biosynthetic process, apoptosis, ribonucleoprotein complex assembly, and M T O R C 1 signaling. Panel E shows immunofluorescence staining of white adipose tissue for M a t 2 a and E r g in wildtype and M y c t 1 e c K O mice, with yellow arrowheads indicating positive staining. Panel F provides a quantification graph of the data shown in Panel E, indicating a significant increase in M a t 2 a slash E r g plus, nuclei in M y c t 1 e c K O mice.
Ablation of Myct1 upregulates pathways involved in protein folding, ribonucleoprotein assembly, and mTORC1 signaling. (A) Experimental workflow of the scRNA-seq experiment. CD45negCD31+ cells were sorted from mesenteric fat of 12-wk-old wild-type and Myct1ecKO mice. n = 2 per genotype. (B) UMAP representation of cells retained in the scRNA-seq and colored according to cell type. BECs, blood endothelial cells; LECs, lymphatic endothelial cells; vSMCs, vascular smooth muscle cells. (C)Myct1 is efficiently deleted from ECs in Myct1ecKO mice. Distribution of Myct1, Pecam1, and Cdh5 expression (ln[normalized counts +1]) in the BEC cluster in wild-type and Myct1ecKO cells. (D) GO terms and Hallmark gene sets overrepresented among upregulated (red bars) or downregulated (gray bars) genes in Myct1ecKO BECs compared with wild-type BECs. (E)Myct1 deficiency in ECs increases protein levels of mTORC1 target Mat2a (methionine adenosyltransferase 2A). Staining of WAT of Myct1ecKO or wild-type control mice for Mat2a (green) and pan-endothelial marker Erg (magenta). Scale bar, 50 µm. (F) Quantification of data shown in E. n = 8 mice per genotype from three independent experiments. mean ± SD; paired t test, P = 0.0191 (*). Icons used in A were created with BioRender.com and modified in Affinity. See also Fig. S3, A–D.