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. 2020 Sep 6;9(9):2037. doi: 10.3390/cells9092037

Figure 3.

Figure 3

Involvement of EAATs in the glutamate neuroprotection against α-syn plus Rot toxicity. (A,C) Intracellular ATP content (A) and extracellular LDH (C) after 24 h exposure to α-syn (10 nM) plus Rot (300 nM) and 23 h exposure to α-syn (10 nM) plus Rot (300 nM) followed by 1 h exposure to glutamate (500 µM) in the presence or in the absence of DL-threo-beta-benzyloxyaspartate (DL-TBOA; 300 µM). (B,D) After 48 h of EAAT3 silencing, ATP levels (B) and cell damage (D) were evaluated after 24 h exposure to α-syn (10 nM) plus Rot (300 nM) and 23 h exposure to α-syn (10 nM) plus Rot (300 nM) followed by 1 h exposure to glutamate (500 µM). Statistical differences were assessed by one-way ANOVA followed by Dunnet’s post hoc test. (A) F (4,15) = 13.75. Each column represents the mean ± S.E.M. of n = 4 independent experiments performed in triplicate. * Significant versus Ctl (p < 0.01) and α-syn+Rot+Glut (p < 0.05); ** significant versus α-syn+Rot (p < 0.05), α-syn+Rot+Glut+DL-TBOA and α-syn+Rot+DL-TBOA (p < 0.01); # significant versus Ctl (p < 0.001) and α-syn+Rot+Glut (p < 0.01); ## significant versus Ctl (p < 0.0001) and α-syn+Rot+Glut (p < 0.01). (B) F (4,15) = 12.25. Each column represents the mean ± S.E.M. of n = 4 independent experiments performed in triplicate. * Significant versus Ctl and α-syn+Rot+Glut (p < 0.001); ** significant versus α-syn+Rot, α-syn+Rot+Glut+siEAAT3 and α-syn+Rot+siEAAT3 (p < 0.001); # significant versus Ctl (p < 0.0001) and α-syn+Rot+Glut (p < 0.001). (C) F (4,25) = 11.58. Each column represents the mean ± S.E.M. of n = 6 independent experiments performed in triplicate. * Significant versus Ctl (p < 0.0001) and α-syn+Rot+Glut (p < 0.05); ** significant versus all groups (p < 0.05). (D) F (4,10) = 20.73. Each column represents the mean ± S.E.M. of n = 3 independent experiments performed in triplicate. * Significant versus Ctl (p < 0.001) and α-syn+Rot+Glut (p < 0.05); ** Significant versus all groups (p < 0.05); # Significant versus Ctl (p < 0.0001) and α-syn+Rot+Glut (p < 0.05). siEAAT3 = siRNA for EAAT3.