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. 2020 Jun 18;11(6):469. doi: 10.1038/s41419-020-2663-1

Fig. 3. Involvement of SMCT1 in inhibitory effect of NaB on high cholesterol-induced ROS generation, BACE1 expression, and Aβ accumulation.

Fig. 3

a SK-N-MC cells were pretreated with NaB and ibuprofen (500 µM) for 30 min prior to treatment of high cholesterol for 48 h where DCF-DA was detected by luminometer. n = 4. *p < 0.05 versus control, #p < 0.05 versus high cholesterol treatment, $p < 0.05 versus treatment of high cholesterol and NaB. b Cells were pretreated with NaB and PTX (200 nM) for 30 min prior to treatment of high cholesterol for 48 h where DCF-DA was detected by luminometer. *p < 0.05 versus control, #p < 0.05 versus high cholesterol treatment. c Cells were pretreated with NaB and ibuprofen for 30 min prior to treatment of high cholesterol for 72 h where ROS with DCF-DA were measured by flowcytometer. Total cell counts = 1.0 × 104 cells. n = 4. *p < 0.05 versus control, #p < 0.05 versus high cholesterol treatment, $p < 0.05 versus treatment of high cholesterol and NaB. d Cells were pretreated with NaB and PTX for 30 min prior to treatment of high cholesterol for 72 h where ROS with DCF-DA were measured by flowcytometer. Total cell counts = 1.0 × 104 cells. n = 4. *p < 0.05 versus control, #p < 0.05 versus high cholesterol treatment. e Cells were transfected with SMCT1 siRNA or NT siRNA for 12 h, and pretreated with NaB for 30 min prior to treatment of high cholesterol for 72 h where ROS with DCF-DA were measured by flowcytometer. Total cell counts = 1.0 × 104 cells. Data are presented as a mean ± S.E.M. n = 3. *p < 0.05 versus control with NT siRNA transfection, #p < 0.05 versus high cholesterol treatment with NT siRNA transfection, $p < 0.05 versus treatment of high cholesterol and NaB with NT siRNA transfection. f Cells were pretreated with NaB and ibuprofen for 30 min prior to treatment of high cholesterol for 24 h. The expression levels of BACE1 were analyzed by western blot. β-actin was used as a loading control. n = 3. *p < 0.05 versus control, #p < 0.05 versus high cholesterol treatment, $p < 0.05 versus treatment of high cholesterol and NaB. g Cells were pretreated with NaB and PTX for 30 min prior to treatment of high cholesterol for 24 h. The expression levels of BACE1 were analyzed by western blot. β-actin was used as a loading control. n = 3. *p < 0.05 versus control, #p < 0.05 versus high-cholesterol treatment. h Cells were transfected with SMCT1 siRNA or NT siRNA for 12 h, and pretreated with NaB for 30 min prior to treatment of high cholesterol for 24 h. The expression levels of BACE1 were analyzed by western blot. β-actin was used as a loading control. n = 3. *p < 0.05 versus control with NT siRNA transfection, #p < 0.05 versus high cholesterol treatment with NT siRNA transfection, $p < 0.05 versus treatment of high cholesterol and NaB with NT siRNA transfection. i Cells were transfected with SMCT1 siRNA or NT siRNA for 12 h, and pretreated with NaB for 30 min prior to treatment of high cholesterol for 72 h. Aβ concentration of medium samples was detected by using ELISA kit. Data are presented as a mean ± S.E.M. n = 4. *p < 0.05 versus control with NT siRNA transfection, #p < 0.05 versus high cholesterol treatment with NT siRNA transfection, $p < 0.05 versus treatment of high cholesterol and NaB with NT siRNA transfection. All blot images shown are representative.