A. Expression of TIM-3 and DBA on uNK cells during gestation.

<p>A steady and similar increase in the expression of both TIM-3 and DBA is observed between GD 7.5 and 12.5, the time period when the uNK cells are most abundant in the uterus. TIM-3 and DBA expression were determined on the CD3^- CD122⁺ NK1.1^- DX5^- cells isolated from the uterus of pregnant CBA mice allogeneically mated with C57BL/6 males (n = 8). Data are presented as mean± SEM and are representative of at least 3 experiments. Data show the percentage of CD3- CD122+ NK1.1- DX5- cells expressing DBA and TIM-3 on different GDs. Statistical analysis was performed using 2 way ANOVA. B. Effect of TIM-3 blockade on uNK cell population size. Percentage of CD3^- CD122⁺ NK1.1^- and DX5^- cell population in control and RMT3-23 treated pregnant CBA mice at GD 7.5, 10.5 and 12.5. There was no significant change in the number of uNK cells between control and treated groups. Data are presented as mean± SEM and are representative of at least 3 experiments. Statistical analysis was done by one-way analysis of variance (ANOVA) using Kruskal-Wallis test. C. Effect of TIM-3 blockade on uNK cell cytotoxicity. No significant change was observed in the cytotoxicity of uNK cells following treatment with RMT3-23. Data are presented as mean± SEM and are representative of at least 3 experiments. Statistical analysis was done using unpaired t test. D. Effect of TIM-3 blockade on Granzyme production by uNK cells. A small but significant increase in the production of Granzyme B was observed in the RMT3-23 treated uNK cells in comparison to control. Data are presented as mean± SEM and are representative of at least 3 experiments. Statistical analysis was done using unpaired t test.