Figure 9
From: Defective Sphingosine-1-phosphate metabolism is a druggable target in Huntington’s disease
Pharmacological modulation of S1P metabolizing enzyme activity regulates cell survival in HD striatal derived cells. Cropped immunoblotting of SGPL1 protein (A) and mRNA expression levels (B) from STHdh7/7 and STHdh111/111 cells. Values are mean ± SD of three experiments performed in triplicate. *p < 0.05 (Unpaired t-test). Analysis of apoptosis in STHdh cell lines first cultured for three days in complete medium in the absence and presence of 5 μM THI and then incubated for five hours in serum free medium to induce apoptosis (C). Values are mean ± SD of two experiments performed in quadruplicate. **p < 0.001(Unpaired t-test). Schematic representation of the pharmacological interventions to modulate the activity of S1P-metabolizing enzymes (D). Cropped immunoblotting of SPHK2 protein (E) and mRNA expression levels (F) from STHdh7/7 and STHdh111/111 cells. Values are mean ± SD of three experiments performed in triplicate. *p < 0.05 (Unpaired t-test). Analysis of apoptosis in STHdh cell lines incubated for five hours in serum free media in the presence or absence of either 1.5 μM K145 (G) or 5 and 10 μM EMD567731 (H) SPHK2 inhibitors. Values are mean ± SD of three experiments, each performed in triplicate. *p < 0.05; **p < 0.001 (Unpaired t-test). Cropped immunoblotting of SPHK1 protein (I) and mRNA expression levels (L) from STHdh7/7 and STHdh111/111 cells. Values are mean ± SD of three experiments performed in triplicate. *p < 0.05 (Unpaired t-test). Analysis of apoptosis in STHdh cell lines incubated for five hours in serum free media in the presence or absence of 5 μM SK1-I (2R, 3S, 4E)-N-methyl-5-(4′-pentylphenyl)-2-aminopent-4-ene-1,3-diol) (M) and 15 μM DMS (N,N-Dimethylsphingosine) (N). Data are mean ± SD of three experiments, each performed in triplicate. **p < 0.001 (Unpaired t-test). Apoptosis in STHdh cell lines incubated for five hours in serum-free medium in the presence or absence of 50 μM K6PC-5 (O). Data are mean ± SD of three experiments, each performed in triplicate. **p < 0.001; ****p < 0.0001 (Unpaired t-test). Cropped immunoblotting of AKT and ERK phosphorylation in cellular protein extracts from STHdh7/7 (P) and STHdh111/111 (Q) cell lines measured at different time points after K6PC-5 treatment.
