Estrogen regulation of lupus-associated miRNAs expression in splenocytes from orchidectomized NZB/W
mice. (A) Real-time RT-PCR analysis of miRNA expression in splenocytes from 10-wk-old and 32-wk-old intact male, placebo- and estrogen-treated orchidectomized male NZB/WF1 mice. The expression level of a specific miRNA in all other groups was shown as relative level to the 10-wk-old intact male mice. The graphs represent the means ± SEMs (n = 4 each for 10-wk-old intact and placebo-treated mice groups, n = 5 each for 10-wk-old estrogen-treated, 32-wk-old intact, and 32-wk-old placebo-treated mice groups, and n = 2 for 32-wk-old estrogen-treated mice group). One-way ANOVA with Tukey-Kramer all pair's comparison tests were performed to compare the expression among the age-matched intact, placebo-, and estrogen-treated mice. Asterisk indicates the statistical significance between the age-matched placebo control and estrogen-treated mice, and the number sign indicates the statistical significance between the age-matched intact control and estrogen-treated mice. Asterisk and number sign depict p < 0.05; double asterisk and double number sign depict p < 0.01. (B) Anti-dsDNA autoantibody ELISA. The figure shows the serum anti-DNA autoantibody levels in NZB/WF1 male mice from aforementioned six treatment groups. Four C57BL6/ mice were included as negative controls. The bold black line represents the mean serum autoantibody value of each group.