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

Figure 4.

Figure 4

Involvement of NCX 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 SN-6 (1 µM). (B,D) After 48 h of NCX1 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). The experiments reported in panel D were conducted simultaneously with the experiments reported in panel 3D. Statistical differences were assessed by one-way ANOVA followed by Dunnet’s post hoc test. (A) F (4,30) = 5.737. Each column represents the mean ± S.E.M. of n = 7 independent experiments performed in triplicate. * Significant versus Ctl (p < 0.001) and α-syn+Rot+Glut (p < 0.01); ** significant versus α-syn+Rot, α-syn+Rot+Glut+SN-6 and α-syn+Rot+SN-6 (p < 0.01); # significant versus Ctl and α-syn+Rot+Glut (p < 0.01). (B) F (4,20) = 13.62. Each column represents the mean ± S.E.M. of n = 5 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+siNCX1 (p < 0.001) and α-syn+Rot+siNCX1 (p < 0.05); # significant versus Ctl and α-syn+Rot+Glut (p < 0.05). (C) F (4,15) = 17.71. Each column represents the mean ± S.E.M. of n = 4 independent experiments performed in triplicate. * Significant versus Ctl (p < 0.0001) and α-syn+Rot+Glut (p < 0.01); ** significant versus all groups (p < 0.05 versus Ctl and α-syn+Rot+Glut+SN-6, p < 0.01 versus α-syn+Rot and α-syn+Rot+SN-6); # significant versus Ctl (p < 0.0001) and α-syn+Rot+Glut (p < 0.05). (D) F (4,15) = 19.99. Each column represents the mean ±  S.E.M. of n = 4 independent experiments performed in triplicate. * Significant versus Ctl (p < 0.0001) and α-syn+Rot+Glut (p < 0.01); ** significant versus all groups (p < 0.05 versus Ctl and α-syn+Rot+Glut+siNCX1, p < 0.01 versus α-syn+Rot and α-syn+Rot+siNCX1); # significant versus Ctl (p < 0.0001) and α-syn+Rot+Glut (p < 0.05). siNCX1 = siRNA for NCX1.