FIGURE LEGENDS
Figure 1: Baseline expression of monocyte CD11b, TLR4, ER-α, and ER-β in neonates and adults. PBMCs were isolated from adult peripheral venous blood and umbilical cord blood from neonates and analysed by flow cytometry. No statistically significant differences in (A) CD11b (B) TLR4 (C) ER-α or (D) ER-β monocyte receptor expression were observed between adults versus neonates or males versus females. Males = white, black = females. n = 10 per group. Data expressed as percentage of total PBMCs. Statistical analysis by ANOVA with Bonferroni correction for multiple comparisons. PBMCs, peripheral blood mononuclear cells.
Figure 2: Treatment with E2, ICI 182,780, or LPS individually or in combination do not modulate monocyte CD11b or TLR4 in neonates or adults. PBMCs were isolated from adult peripheral venous blood and umbilical cord blood from neonates and stimulated for 1 hour at 37°C with vehicle, E2 (10 nM), ICI 182,780 (10 nM) and LPS (1 μg/mL) individually or in combination and analysed by flow cytometry. No statistically significant differences in neonatal (A) or adult (B) CD11b or neonatal (C) or adult (D) TLR4 monocyte receptor expression were observed following treatment or between males versus females. Males = white, black = females. n = 10 per group. Data expressed as percentage of total PBMCs. Statistical analysis by ANOVA with Bonferroni correction for multiple comparisons. PBMCs, peripheral blood mononuclear cells; LPS, lipopolysaccharide; E2, 17-β-estradiol.
Figure 3: Sex-specific differences in adult monocyte ER-β expression in response to ICI 182,780. PBMCs were isolated from adult peripheral venous blood and umbilical cord blood from neonates and stimulated for 1 hour at 37°C with E2 (10 nM), ICI 182,780 (10 nM) and LPS (1 μg/mL) individually or in combination and analysed by flow cytometry. No statistically significant differences in neonatal (A) or adult (B) ER-α or neonatal (C) ER-β monocyte receptor expression were observed following treatment or between males versus females. A statistically significant difference between male versus female adults was observed in ER-β monocyte receptor expression following ICI 182,780 treatment (*p<0.05). Males = white, black = females. n = 10 per group. Data expressed as percentage of total PBMCs. Statistical analysis by ANOVA with Bonferroni correction for multiple comparisons. PBMCs, peripheral blood mononuclear cells; LPS, lipopolysaccharide; E2, 17-β-estradiol.
Figure 4: E2 attenuates LPS-induced ROS in female neonates.PBMCs were isolated from adult peripheral venous blood and umbilical cord blood from neonates and stimulated for 1 hour at 37°C with E2 (10 nM) and LPS (1 μg/mL) individually or in combination and analysed by flow cytometry. LPS increases ROS production in (A) neonates and (B) adults and this effect is not sex-specific. E2 reduces the LPS effect in both neonates and adults. Female neonates exhibited significantly less ROS following E2 and LPS treatment compared to male neonates (*p<0.05). Males = white, black = females. n = 10 per group. Data expressed as percentage of total PBMCs. Statistical analysis by ANOVA with Bonferroni correction for multiple comparisons. MFI, mean fluorescence intensity; LPS, lipopolysaccharide; E2, 17-β-estradiol.