Fig 1: T-cell development in Gli3+/+, Gli3+/- and Gli3-/- E17.5 plus 4 days in FTOC and E18.5 thymus. Flow cytometry analysis. Scatter plots show mean±s.e.m. Each point represents thymus from an individual mouse embryo. (A-H) E17.5 FTOC for 4 days. Gli3+/+ (n=14), Gli3+/- (n=14) and Gli3-/- (n=11). (I,J) E18.5 Gli3+/+ (n=3) and Gli3-/- (n=3) thymus. (A) CD8 against CD4. (B) Percentage of populations (relative to mean of WT) giving significance by Student's t-test compared with WT for SP4 (Gli3+/-, P<0.03; Gli3-/-, P<0.001), SP8 (Gli3-/-, P<0.03) and SP4:SP8 ratio (Gli3+/-, P<0.02; Gli3-/-, P<0.0006). (C) CD3 staining on Gli3+/+, Gli3+/- and Gli3-/- thymocytes, giving the percentage of CD3hi cells. (D) Percentage of CD3hi thymocytes in Gli3+/+, Gli3+/- and Gli3-/- (relative to mean of WT littermate). (E) CD8 against CD4, gated on CD3hi. (F) Percentage of populations gated on CD3hi, giving significance by Student's t-test compared with WT for CD3hi SP4 (Gli3+/-, P<0.002; Gli3-/-, P<0.0004), CD3hi SP4:SP8 ratio (Gli3-/-, P<0.002) and CD3hi DP (Gli3+/-, P<0.05; Gli3-/-, P<0.02). (G) HSA against Qa2 expression, gated on SP4 cells from Gli3+/+ and Gli3-/-. (H) Relative percentage of HSA- Qa2+ cells in the SP4 population, giving significance by Student's t-test compared with WT (P<0.002). (I) Percentage of CD69+ cells in SP4 and DP populations, giving significance by Student's t-test for SP4 (P<0.07) and DP (P<0.05). (J) CD69 expression on SP4 and DP cells in a representative experiment.
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