Extended Data Fig. 8: Cellular GGR controls SMC switching.
From: GLS2 links glutamine metabolism and atherosclerosis by remodeling artery walls
(a) Experimental outline. Mouse smooth muscle cell line (MOVAS) was treated for 24 hours with glutaminase inhibitors in presence or absence of glutamate supplementation to set up GGRhigh and GGRlow conditions (upper panel). Venn diagram highlights genes specifically regulated by GGRhigh and GGRlow conditions and those oppositely regulated between these conditions. (b) RNA-seq results on genes belonging to Gln- or Glu-dependent metabolism (upper panel) (n = 3 independent cultures; log2 fold change was used to assess statistical significance with two-tailed t test, with P < 0.05 indicated by an asterisk). Schematic representation of upregulated (red) and downregulated (blue) Gln- and Glu-dependent transcripts in SMC in GGRhigh and GGRlow conditions. (c) Differentially expressed genes between these cell culture conditions were subjected to gene set enrichment analysis (GSEA). The top 4 downregulated or oppositely regulated pathways are shown. (d) Immunoblot and quantification of Glu-tubulin cultured on soft (1 kPa) or stiff (50 kPa) conditions with the indicated conditions (n = 3 independent cultures, P values displayed; two-tailed t test). (e) Representative images (upper panel) and quantification of gel contraction (lower right panel) (n = 12 replicates with four independent cultures, P values displayed; two-tailed t test). Quantification of force magnitude using traction force microscopy (lower right panel) (n = 9 replicates with three independent cultures, P values displayed; two-tailed t test). Data from individual cultures and results are means ± s.e.m. Two-tailed Student’s t-tests were used.
