Spleen iNKT cell number is not affected by E2 treatment. Sham-operated (▪) and either placebo- (□) or E2-treated (▨) ovariectomized (ovx) female mice were killed at 8 weeks of age. Splenocytes were labeled with fluorochrome-conjugated anti–αβ-TCR and anti-NK.1.1 Abs and with either CD1d/α-GalCer-tetramer or CD1d/vehicle-tetramer, then analyzed by flow cytometry. Absolute number of conventional T (αβ-TCR⁺ NK.1.1^-) lymphocytes (A) and iNKT cells— namely, αβ-TCR⁺ NK.1.1⁺ (B) and αβ-TCR⁺ CD1d/α-GalCer-tetramer positive cells (C)—are given as means ± SEMs from 16 individual mice per group. In each group, control staining with CD1d/vehicle-tetramer was less than 0.05%. ^*P < .05; ^**P < 01. NS indicates not significant.