Fig 2

Rag2:EGFP-CA-CaMKII is expressed in thymic T cells and immature kidney marrow B lymphocytes.

(A-F) Rag2:EGFP-CA-CaMKII colocalizes with rag2:DsRed in the thymus at (A-C, confocal images A’-C’) 4 dpf and (D-F) 5 mpf but does not colocalize in the skin at (skin magnified D’-F’) 5 mpf fish. (G) FACS analysis of kidney marrow cells from rag2:EGFP-CA-CaMKII; rag2:DsRed fish at 3 mpf. Gated populations: erythrocytes (red gate), lymphocytes (green gate), precursors (pink gate), and myelomonocytes (blue gate). (H) The EGFP and DsRed positive population for each sample is predominantly found in the lymphocyte gate, as expected. (I) Lymphocytes were further defined as double negative (DN), single positive EGFP (gSP) single positive DsRed (rSP), and double positive (DP). Rag2:EGFP-CA-CaMKII; rag2:DsRed kidney marrow lymphocyte cells were sorted at (J,K) 6 mpf and (L,M) 12 mpf then analyzed for B cell (rag2, rag1, igiv1s1, cd79a, ighm-C, ight-C, igic1s1) and T cell (lck), marker gene expression using RT-PCR (K,M). Ef1a served as a control. Scale bars: 100 μm in C and 500 μm in F.