Fig. 1

The expression of characteristic glial cell proteins, cell migration, intracellular α-KG, and histone demethylation of BMSCs were increased in homoglutamate microenvironment. Glutamate (2 mM) was incubated with BMSCs for 24 h. A and B Representative immunoblot bands and histogram of relative expression for the GFAP (Control vs. Glu: 1 vs. 1.63 ± 0.23, P < 0.05), S100B (Control vs. Glu: 1 vs. 1.32 ± 0.03, P < 0.01), and GDNF (Control vs. Glu:1 vs. 1.64 ± 0.22, P < 0.05) proteins. GAPDH was used as a loading control. C and D Transwell migration experiment of BMSCs and statistical analysis (Control vs. Glu: 140 ± 20.3 vs. 256 ± 27.4, P < 0.01). E The transcripts of Glud1 were determined by a reverse transcription-polymerase chain reaction (RT-PCR) assay (Control vs. Glu: 1.0 ± 0.18 vs. 1.5 ± 0.22, P < 0.05). F–H The intracellular α-KG (Control vs. Glu: 3.34 ± 0.18 nmol vs. 4.38 ± 0.37 nmol, P < 0.01), succinate (Control vs. Glu: 24.7 ± 2.6 nmol vs. 18.7 ± 23 nmol, P < 0.05), concentration and α-kg/succinate ratio (Control vs. Glu: 0.13 ± 0.01 vs. 0.24 ± 0.05, P < 0.01) of BMSCs. I and J Representative immunoblot bands and histogram of relative expression for the H3K9me3 (Control vs. Glu: 1 vs. 0.42 ± 0.09, P < 0.01), H3K9me1 (Control vs. Glu: 1 vs. 1.90 ± 0.37, P < 0.05), H3K27me3 (Control vs. Glu: 1 vs. 0.60 ± 0.17, P < 0.05), and H3K27me1 (Control vs. Glu: 1 vs. 1.7 ± 0.14, P < 0.01) proteins. H3 was used as a loading control. Glu: glutamate; α-KG: alpha-ketoglutarate; Glud1: glutamate dehydrogenase 1. These results are representative of at least three times independent experiments. *P < 0.05, **P < 0.01