Supplementary Figure 7: FTY720 does not affect performance of SCID mice in the Morris water maze, marble burying, exploratory activity and cued fear conditioning test.

SCID mice were treated orally each day by gavage with saline or FTY720 (1 mg/kg) starting 16 h prior to the (a-c) MWM paradigm, (d-f) behavioral testing, and (g) tone-shock paired conditioned fear testing. (a-c) FTY720 did not affect the performance of SCID mice in MWM. (a) Escape latencies (two-way repeated measures ANOVA; interaction: F(4,60) = 1.43; P = 0.23; day: F(4,60) = 1.96; P =0.11; treatment: F(1,60) = 0.64; P = 0.44) and (b) swim speeds (two-way ANOVA; interaction: F(4,60) = 0.26; P = 0.90; day: F(4,60) = 1.68; P = 0.17; treatment: F(1,60) = 0.33; P = 0.57) of FTY720 and saline treated mice did not significantly differ on the five days of fixed platform training. (c) The percentage of time spent in the in each quadrant (T, target quadrant; L, left quadrant; O, opposite quadrant; R, right quadrant) during a 120 s probe trial. 24 h after the fifth day of training also did not differ in mice treated with FTY720 or saline (two-way repeated measures ANOVA; interaction: F(3,45) = 0.10; P = 0.96; quadrant: F(3,45) = 1.15; P = 0.34; treatment: F(1,45) = 0.93; P = 0.35). T, target quadrant; L, left quadrant; O, opposite quadrant; R, right quadrant. (d) In the marble burying test, SCID mice buried a similar number of marbles regardless of FTY720 treatment (t-test: t (1,18) = 0.63; P = 0.54). (e-f) FTY720 did not affect (e) distance traveled (t-test: t(1,18) = 0.32; P = 0.75) or (f) time thigmotaxic during a 3 min exposure to a novel environment (t-test: t(1,18) = 1.61; P = 0.13). (g) SCID mice treated with FTY720 or saline displayed similar freezing and extinction in a tone-shock paired conditioning paradigm. Data represent percent time freezing during the 3 min before tone-shock pairings (preshock), 60 s after tone-shock pairings (post-shock), and during 200 s tone on extinction days. *P < 0.001, vs. pre-shock (Bonferroni post hoc test). ^# P < 0.001, compared to day 2, the first day of extinction (Bonferroni post hoc test). Both treatment groups had similar levels of freezing preshock, post-shock, and on extinction day 1 (two-way ANOVA; interaction: F(2,36) = 0.51; P = 0.61; time: F(2,36) = 146.54; P <0.0001; treatment: F(1,36) = 0.67; P = 0.42) and exhibited similar rates of extinction (two-way repeated measures ANOVA, interaction: F(4,72) = 0.28; P = 0.89; extinction session: F(4,72) = 8.33; P <0.0001; treatment: F(1,72) = 0.23; P = 0.64). Tests in (d-g) were performed in the same cohort of mice. N = 7-10 mice/group. Data are mean ± s.e.m.