Effect of Lipopolysaccharide and TNFα on Neuronal Ascorbic Acid Uptake

Figure 2

Effect of exposure of SH-SY5Y cells and mouse brain with LPS on different aspects of SVCT2 functional expression. (a) SH-SY5Y cells were serum-deprived overnight and exposed to LPS (20 μg/ml). After 48 h, AA uptake was determined. (b) SH-SY5Y cells were exposed to LPS for 48 h, and the protein was prepared to perform western blot analysis to determine the hSVCT2 protein expression levels. (c) SLC23A2 full-length promoter activity was determined in LPS-treated SH-SY5Y cells. (d, f) Total RNA isolated from LPS-administered and control mouse brain were used to determine the mSVCT2 and NLRP3 mRNA expression levels by RT-qPCR. (e) Protein samples prepared from mouse brain tissue of LPS (5 mg/kg body weight; 72 h) exposed and controls were subjected to western blotting to determine mSVCT2 protein expression levels. (g) Total RNA prepared from LPS-injected and control mouse brain were subjected to RT-qPCR to determine the mSVCT2 hnRNA expression levels. Values are of at least 3-5 independent investigations utilizing multiple batches of SH-SY5Y cells or at least 3-5 pairs of mice. ; ; .

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