Fig. 3

osm Inhibits Lymphocyte Priming and Differentiation by Repressing Lymphoid Gene Expression

(A) ISH at 4.5 dpf of rag1 thymus staining with quantification of thymus area, where p = 0.0087. Scale, 50 ?m.

(B) Live imaging of CHT region in ikaros:eGFP embryos at 48 hpf. Each arrow indicates a single ikaros:eGFP^high cell. Scale, 100 ?m.

(C) Quantification of the number of ikaros:eGFP^high cells at 48 hpf, p = 0.0028.

(D) FACS sorting and analysis of ikaros:eGFP^high cells at 48 hpf (p = 0.0112). Values indicated on FACS plots are mean ± SEM, graphs are means ± SD.

(E) Quantification of ikaros:eGFP^high cells.

(F?I) qPCR analysis of il7r (F) (p = 0.0017), irf4a (G) (p = 0.0001), ccr9a (H) (p = 0.1265), and ikaros (I) (p = 0.8030) expression in ikaros:eGFP^high cells FACS sorted at 48 hpf. qPCR data shown are the mean ± SEM of three data points, calculated from three independent experiments. Each separate experiment was conducted in biological triplicates, then averaged to give a single value. NI, non-injected control. +osm, osm full-length mRNA-injected embryos. Statistical analysis was completed using an unpaired, two-tailed Student's t test. ^???p < 0.001; ^??p < 0.01; ^?p < 0.05; NS, p > 0.05.