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Fig. 1 | Biological Research

Fig. 1

From: Thiamine deficiency contributes to synapse and neural circuit defects

Fig. 1

Thiamine deficiency induced spine density decrease in vitro. a TPK mRNA level was significantly reduced in TPK RNAi oligo transfected N2A cell lines (N = 3 culture preparations). b TKP expression examined by western blot in N2A cell lines. TPK overexpression was significantly reduced by RNAi, demonstrating efficiency of the RNAi at the protein level. c TPK mRNA level in the LV-TPK RNAi-GFP infected cultured neurons was significantly lower at 7 days and 14 days after infection, as compared with neurons infected with LV-Ctrl-GFP (N = 4 & N = 3 culture preparations). d Lentiviral-mediated TPK RNAi significantly reduced TDP levels, but not that of thiamine monophosphate (TMP) and thiamine in cultured hippocampal neurons (N = 4 culture preparations). e, f Representative images of spines in cultured hippocampal neurons. Scale bar represents 5 µm. Quantitation of spine density in Ctrl, TPK RNAi and TPK RNAi + human TPK overexpression (hTPK Ovp) neurons (N = 3 culture preparations, n = 28, 26, 27 cells numbers, respectively). TPK knockdown significantly reduced the densities of dendritic spines in cultured hippocampal neurons (P < 0.05) and hTPK overexpression significantly rescued the effect of TPK knockdown on spine density (P < 0.01). All the data are expressed as mean ± SEM from three or four independent experiments. *P < 0.05, **P < 0.001, ***P < 0.0001, student’s t test and one-way ANOVA were used to determine the statistical significance of the differences

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