FIGURE 5.

Decreased P2X1 protein level but not mRNA in Entpd1^−/− vas deferens. A, Western blot analysis showing that P2X1 immunoreactivity is about 3-fold lower in Entpd1^−/− vas deferens, whereas NTPDase2 and P2X4 expression are equivalent in mutant and WT tissues. Error bars show mean ± S.E. of four independent experiments. ***, p < 0.0005. B, immunofluorescence quantification of P2X1 receptor expression in vas deferens (n = 4). *, p < 0.05). C, reverse transcription followed by quantitative PCR analysis shows that P2X1 and P2X4 relative mRNA levels were unchanged between mutant and WT tissues. Expression was normalized to three housekeeping genes (Hprt, Gusb, and Gapdh) and expressed as relative expression = exp (Ct housekeeping − Ct target gene).