Journal: Journal of Cellular Biochemistry

Article Title: Orexin‐A Inhibits Lipopolysaccharide‐Induced Cell Migration in Cultured Mouse Astrocytes via Activation of Orexin 1 Receptor: Involvement of GABA Signaling

doi: 10.1002/jcb.70089

Figure Lengend Snippet: Effect of the treatment of OXA (10 nM) with or without the OX1RA, SB334867 (100 nM) pretreatment on LPS (100 ng/mL)‐induced changes in astrocyte migration. (a) Cell migration assay was performed and the migration images were captured at 0 and 48 h after the start of LPS treatment. (b) A graph representing the percentage of the migrated area, with 0 h set as 0%. The LPS‐induced increase in astrocyte migration was reduced by OXA treatment, and this reduction was restored by pretreatment of OX1RA. Scale bar = 200 μm. Data are expressed as mean ± SEM. n = 7. ** p < 0.01 versus control; ## p < 0.01 and ### p < 0.001 versus LPS + OXA.

Article Snippet: The cells were then pretreated with or without LPS (100 ng/mL) in MEM medium for 30 min, followed by treatment with the OX1R antagonist (OX1RA), SB334867 (100 nM, cat#1960, Tocris Bioscience) for another 30 min.

Techniques: Migration, Cell Migration Assay, Control

Journal: Journal of Cellular Biochemistry

Article Title: Orexin‐A Inhibits Lipopolysaccharide‐Induced Cell Migration in Cultured Mouse Astrocytes via Activation of Orexin 1 Receptor: Involvement of GABA Signaling

doi: 10.1002/jcb.70089

Figure Lengend Snippet: Effect of the treatment of OXA (10 nM) with or without the OX1RA, SB334867 (100 nM) pretreatment on LPS (100 ng/mL)‐induced changes in KCC2 phosphorylation in primary cultured astrocytes. (a and b) Representative Western blots (a) and a graph (b) showed that the LPS‐induced increase in the ratio of phosphorylation (p‐KCC2) at residue Thr‐1007 to total protein expression of KCC2 was reduced by OXA treatment, and this reduction was restored by pretreatment of OX1RA. Data are expressed as mean ± SEM. n = 8. * p < 0.05 versus control; # p < 0.05 versus LPS + OXA.

Article Snippet: The cells were then pretreated with or without LPS (100 ng/mL) in MEM medium for 30 min, followed by treatment with the OX1R antagonist (OX1RA), SB334867 (100 nM, cat#1960, Tocris Bioscience) for another 30 min.

Techniques: Phospho-proteomics, Cell Culture, Western Blot, Residue, Expressing, Control

Journal: Journal of Cellular Biochemistry

Article Title: Orexin‐A Inhibits Lipopolysaccharide‐Induced Cell Migration in Cultured Mouse Astrocytes via Activation of Orexin 1 Receptor: Involvement of GABA Signaling

doi: 10.1002/jcb.70089

Figure Lengend Snippet: Effect of the treatment of OXA (10 nM) with or without the OX1RA, SB334867 (100 nM) pretreatment on LPS (100 ng/mL)‐induced changes in GABA‐immunoreactivity in primary cultured astrocytes. (a and b) Representative images (a) and a graph (b) show that LPS treatment increased GABA immunoreactivity (red) in cultured astrocytes, and OXA treatment further enhanced this increase. This effect was inhibited by pretreatment of OX1RA. Scale bar = 50 μm. Data are expressed as mean ± SEM. n = 3. ** p < 0.01, *** p < 0.0001 versus Control; # p < 0.05, ## p < 0.01 versus LPS + OXA.

Article Snippet: The cells were then pretreated with or without LPS (100 ng/mL) in MEM medium for 30 min, followed by treatment with the OX1R antagonist (OX1RA), SB334867 (100 nM, cat#1960, Tocris Bioscience) for another 30 min.

Techniques: Cell Culture, Control

Journal: Journal of Cellular Biochemistry

Article Title: Orexin‐A Inhibits Lipopolysaccharide‐Induced Cell Migration in Cultured Mouse Astrocytes via Activation of Orexin 1 Receptor: Involvement of GABA Signaling

doi: 10.1002/jcb.70089

Figure Lengend Snippet: Effect of the treatment of OXA (10 nM) with or without the OX1RA, SB334867 (100 nM) pretreatment on LPS (100 ng/mL)‐induced changes in the expression of GAT1 and GAT3 in primary cultured astrocytes. (a–d) Immunocytochemistry (a and b; red) and Western blot analysis (c and d) using antibodies against GAT1 show that LPS treatment reduced GAT1 expression in astrocytes. OXA treatment prevented the LPS‐induced decrease in GAT1 expression, and this effect was blocked by pretreatment of OX1RA. (e–h) Immunocytochemistry (e and f; red) and Western blot analysis (g and h) using antibodies against GAT3 show that LPS treatment reduced GAT3 expression in astrocytes. OXA treatment prevented the LPS‐induced decrease in GAT3 expression, and this effect was also blocked by pretreatment of OX1RA. Scale bar = 50 μm. Data are expressed as mean ± SEM. n = 3‐4. *** p < 0.0001 versus Control; ### p < 0.001 versus LPS + OXA.

Article Snippet: The cells were then pretreated with or without LPS (100 ng/mL) in MEM medium for 30 min, followed by treatment with the OX1R antagonist (OX1RA), SB334867 (100 nM, cat#1960, Tocris Bioscience) for another 30 min.

Techniques: Expressing, Cell Culture, Immunocytochemistry, Western Blot, Control

Journal: Journal of Cellular Biochemistry

Article Title: Orexin‐A Inhibits Lipopolysaccharide‐Induced Cell Migration in Cultured Mouse Astrocytes via Activation of Orexin 1 Receptor: Involvement of GABA Signaling

doi: 10.1002/jcb.70089

Figure Lengend Snippet: Effect of the treatment of OXA (10 nM) with or without the OX1RA, SB334867 (100 nM) pretreatment on LPS (100 ng/mL)‐induced changes in ERK and p38 MAPK phosphorylation in primary cultured astrocytes. (a and b) Representative western blots and graphs showed that the LPS‐induced increases in the ratio of phosphorylation to total protein expression of ERK (a) and p38 MAPK (b) were reduced by OXA treatment, and this reduction was blocked by pretreatment of OX1RA. Data are expressed as mean ± SEM. n = 4–5. * p < 0.05, ** p < 0.01, *** p < 0.001 versus control; # p < 0.05 versus LPS + OXA.

Article Snippet: The cells were then pretreated with or without LPS (100 ng/mL) in MEM medium for 30 min, followed by treatment with the OX1R antagonist (OX1RA), SB334867 (100 nM, cat#1960, Tocris Bioscience) for another 30 min.

Techniques: Phospho-proteomics, Cell Culture, Western Blot, Expressing, Control