Fig. 2
From: Methionine regulates self-renewal, pluripotency, and cell death of GIC through cholesterol—rRNA axis
A Microarray analysis using Metascape. Heat map showing commonly altered pathways in the control and methionine-depleted states of three GICs (MZGC1, MZGC2, and MZGC3). Statistically enriched terms (gene ontology/Kyoto Encyclopedia of Genes and Genomes terms, canonical pathways, hall mark gene sets) are shown with their P value. The 20 families of pathways with the lowest -log10 adjusted P value are depicted. B Methionine depletion decreased (SAM/SAH ratio, which was measured using an ELISA Combo Kit in MZGC1 and MZGC2. C Methionine depletion increased MAT2A and MAT2B mRNA expression in time-dependent manners. Expression of MATII, alpha (MAT2A) and beta (MAT2B), was measured by qPCR after methionine depletion for 6 h, 24 h (day 1), 72 h (day 3), and 144 h (day 6) in MZGC3. D Heat map showing average interested gene mRNA expression level from expression array of MZGC1, MZGC2, and MZGC3 using Prism 7. Methionine depletion for 5 days downregulated TET1, TET2, DNMT3A, and DNMT3B mRNA expression. E: GSEA showing that methionine depletion significantly activated hallmark TNFA signaling via the NFKB pathway. GSEA of differential expression between control and methionine depletion states demonstrated that methionine-depleted GICs were enriched in hallmark TNFA signaling via the NFKB pathway. Heat map of top 20 genes from GSEA analysis is shown on the right. Red column corresponds to upregulated genes and blue column corresponds to downregulated genes