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. 2023 Nov 23;14:7643. doi: 10.1038/s41467-023-43418-5

Fig. 6. SLC35D3 inhibited the activation of NOTCH1 signaling.

Fig. 6

a KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis results of IngWAT from WT and SAKI mice (n = 3 per group). P values are determined using the clusterProfiler R package’s built-in hypergeometric test, and adjusted P values are calculated using Benjamini–Hochberg correction. b GSEA (gene set enrichment analysis) results showed that the Notch signaling pathway was suppressed in the IngWAT of SAKI mice (n = 3 per group). The P values in GSEA are calculated empirically with permutation test by permuting the gene labels at random according to the GSEA algorithm, and FDR q values (corrected P values) are calculated by Benjamini–Hochberg FDR correction. The NES and FDR q values are shown in the images. NES, normalized enrichment score; FDR, false positive rate. c, e qPCR analysis of Slc35d3 and Notch1-related genes in IngWAT from SAKI mice and WT mice (c), as well as in IngWAT from SAKO mice and Flox mice (e); n = 6 per group. d, f Western blot showing the expression of Notch1-related proteins in IngWAT from SAKI mice and their controls (d), as well as in IngWAT from SAKO mice and their controls (f); n = 3 per group. Right panel, summary of the quantification of three independent experiments. g qPCR detection of NOTCH1 expression levels in HEK293T cells overexpressing SLC35D3; n = 6 per group. h Western blot analysis of NOTCH1 protein in HEK293T cells overexpressing SLC35D3. Right panel, summary of the quantification of three independent experiments. i Western blot analysis of NOTCH1 protein in HEK293T cells treated with CHX (cycloheximide). Right panel, summary of the quantification of three independent experiments. Values are the fold induction of gene expression normalized to the housekeeping gene β-Actin (c–i). Data are expressed as the mean ± SEM. Statistical significance was assessed by unpaired two-sided Student’s t test (c–i). The exact P values are shown in the figure. SAKI Slc35d3 adipocyte-specific knock-in, IngWAT inguinal white adipose tissue. Source data are provided as a Source Data file.