Fig. 6

Knockdown of P2Y4R represses inflammation and neuron death of hippocampal CA region in Tat-injected mice. a IFA was performed to test GFAP and P2Y4R protein levels in the brain tissue from Tat-treated mice. b Calculating the percent of P2YR4 expression in GFAP positive cells. The data are represented as the mean ± S.E.M. *P < 0.05. c Schematic diagram of experimental design in mice. Mice were subjected to LV-sh-Ctrl and LV-sh-P2Y4R3 for 14 days before Tat injection, respectively (n = 12 mice/group). d Representative P2Y4R protein expression was analyzed by Western blotting in hippocampal tissues (n = 6). e Relative mRNA expression of TNFα, IL-6, MCP-1, and IP-10 was measured using qPCR assay (n = 6). The relative expression levels of genes were normalized to the expression of mouse Actin. The data are represented as the mean ± S.E.M. *P < 0.05, **P < 0.01 and ***P < 0.001. f Representative neuronal apoptosis in hippocampal CA1 subfield photomicrographs was TUNEL staining and counterstaining with methyl green (n = 5). Original magnification, × 400. g Quantification of the percent of apoptosis positive neurons. Data were obtained from five independent animals and represented as the mean ± S.E.M. *P < 0.05 versus NC with LV-sh-Ctrl; ^#P < 0.05 versus Tat with LV-sh-P2Y4R3. h Representative cresyl violet-stained sections of the hippocampal tissues. Original magnification, × 400. i Quantification of hippocampal CA1 neuron density. Data were obtained from five independent animals and represented as the mean ± S.E.M. **P < 0.01 versus NC with LV-sh-Ctrl; ^#P < 0.05 versus Tat with LV-sh-P2Y4R3