Fig. 4
The expression of the characteristic glial cell proteins and histone demethylation was significantly increased in the BMSCs (OE-Glud1) than the BMSCs (NC-Glud1). A–C The intracellular α-KG (NC: 3.4 ± 0.9 nmol, NC + Glu: 4.9 ± 0.9 nmol, OE: 6.0 ± 1.3 nmol, OE + Glu: 7.4 ± 1.2 nmol), succinate (NC: 23.2 ± 2.6 nmol, NC + Glu: 16.4 ± 2.2 nmol, OE: 16.6 ± 3.0 nmol, OE + Glu: 15.6 ± 3.0 nmol) concentration and α-kg/succinate ratio (NC: 0.15 ± 0.05, NC + Glu: 0.30 ± 0.06, OE: 0.37 ± 0.09, OE + Glu: 0.49 ± 0.12) of BMSCs in different groups. D and E Representative immunoblot bands and histogram of relative expression of for the GFAP (NC: 1, NC + Glu: 1.40 ± 0.08, OE: 1.55 ± 0.14, OE + Glu: 1.89 ± 0.09), S100B (NC: 1, NC + Glu: 2.08 ± 0.27, OE: 2.31 ± 0.27, OE + Glu: 3.67 ± 0.62), and GDNF (NC: 1, NC + Glu: 1.69 ± 0.06, OE: 1.75 ± 0.17, OE + Glu: 2.26 ± 0.16) in each group. GAPDH was used as a loading control. F–G Representative immunoblot bands and histogram of relative expression of for the H3K9me3 (NC: 1, NC + Glu: 0.54 ± 0.13, OE: 0.57 ± 0.14, OE + Glu: 0.17 ± 0.07), H3K9me1 (NC: 1, NC + Glu: 2.10 ± 0.14, OE: 1.73 ± 0.24, OE + Glu: 2.73 ± 0.38), H3K27me3 (NC: 1, NC + Glu: 0.55 ± 0.09, OE: 0.48 ± 0.09, OE + Glu: 0.23 ± 0.07), and H3K27me1(NC: 1, NC + Glu: 2.09 ± 0.33, OE: 2.24 ± 0.41, OE + Glu: 3.93 ± 0.54) in each group. H3 was used as a loading control. H–I The ELISA level of GDNF (NC: 270 ± 38.98 pg/ml, NC + Glu: 349 ± 13.33 pg/ml, OE: 363.68 ± 36.4 pg/ml, OE + Glu:448.91 ± 13.54 pg/ml) and S100B (NC: 41.05 ± 9.05 pg/ml, NC + Glu: 83.34 ± 6.29 pg/ml, OE: 91.87 ± 4.47 pg/ml, OE + Glu: 111.56 ± 9.77 pg/ml) in the culture medium samples of BMSCs in different groups. J The transcripts of Glud1 (NC: 1 ± 0.13, NC + Glu: 1.39 ± 0.15, OE: 3.52 ± 0.77, OE + Glu: 3.71 ± 0.48) were determined by a RT-PCR assay. Glu: glutamate; Glud1: glutamate dehydrogenase 1; OE: overexpression; NC: negative control. These results are representative of at least three times independent experiments. *P < 0.05, **P < 0.01, NS: no significance