Fig. 3

setdb1b−/− mutation causes a defect in T lymphopoiesis. a qPCR analysis of the relative mRNA levels of setdb1b in coro1a-lyz-, coro1a+lyz- and coro1a+lyz+ cells at 60 hpf, n = 3. b–f WISH analysis of lck, il7r, ikaros and coro1a in thymus (white circles) of setdb1b−/− mutants and WT at 96 hpf. Red, dark gray and light gray squares indicate normal (lck: 30-60; il7r: 30-60; ikaros: 30-60; and coro1a: 45-90), small (lck: 10-30; il7r: 10-30; ikaros: 10-30; and coro1a: 20-45), and absence (lck: 0-10; il7r: 0-10; ikaros: 0-10; and coro1a: 0-20) foci in thymus, respectively. WISH quantifications are shown in (c-f), 3 independent experiments. g qPCR analysis of the relative mRNA levels of ikaros, lck, il7r, tcrα, tcrδ and foxp3a in whole embryos of setdb1b−/− mutants and siblings at 96 hpf, n = 3. h, i Confocal imaging showing lymphocyte in thymus (white circles) of control MO or setdb1b MO-injected WT embryos in Tg(rag2:DsRed) background at 96 hpf. Quantification of rag2+ cells is shown in (i), 3 independent experiments. j, k Representative FACS of rag2:DsRed cells in control MO or setdb1b MO-injected WT embryos at 96 hpf. Quantification of rag2+ cells is shown in (k), n = 5. l Schematic diagram showing the construct of full-length atf7ip gene driven by the coro1a promoter on tol2 vector. m Immunoblot and quantification depicting the protein level of Setdb1 in embryos injected with coro1a:setdb1b/tol2 constructs at 96 hpf, n = 3. n, o WISH analysis of rag1 in thymus (white circles) of WT, setdb1b−/− mutants and embryos injected with coro1a:setdb1b/tol2 constructs at 96 hpf. Red, dark gray and light gray squares indicate normal (45-90), small (10-45), and absence (0-10) foci in thymus, respectively. Quantifications of WISH are shown in (o), 3 independent experiments. Scale bar, 100 μm (b, h, n). Quantifications (a, c–f, g, i, k, m, o) are represented as mean ± SD; ns, P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Student’s t test (unpaired, two-tailed). Source data are provided as a Source Data file.