Fig. 1

Tat upregulated P2Y4R expression in mouse astrocytes. Primary mouse astrocytes were incubated in a serum-free medium overnight followed by treating with soluble Tat protein for various time points. a Representative mRNA levels of P2Y receptors were measured by qPCR assay. These data are from three independent experiments. The relative expression levels of genes were normalized to the expression of mouse Actin. NC negative control. b Representative protein expression of P2Y4R in mouse primary astrocytes was detected by Western blotting assay. c Quantification of P2Y4R protein level. Numbers under the bands were the relative intensity of the bands after calculating for loading control using β-actin. The relative value of proteins in NC group was considered as “1”; same for all of following Western blotting Figures. Error bars represent the mean ± S.E.M. These data are from three independent experiments. **P < 0.01 and ***P < 0.001 versus NC group. d Immunofluorescent assay for GFAP (green) and P2Y4R (red) in primary mouse astrocytes. Scale bars, 50 μm. iTat inactivated Tat, NC negative group. e Western blotting assay verified protein expression of P2Y4R was in mouse primary astrocytes treated with Tat or iTat for 6 h. The data are from three independent experiments. f qPCR assay and g Western blotting assay were performed to assess the expression of GFAP mRNA and protein. The relative expression levels of genes were normalized to the expression of mouse Actin. Error bars represent the mean ± S.E.M. The data are from three independent experiments. *P < 0.05 versus NC group