Fig. 2

Inflammasome Action Regulates HSPC Formation (A) Expression of runx1/cmyb in embryos with or without morpholino (MO)-mediated caspa knockdown or 1% glucose treatment (12?36 hpf) assessed by WISH. (B) Phenotypic distribution plot of runx1/cmyb expression scored in embryos from (A) (n ? 50 /condition). (C) caspa knockdown reduces the frequency of CD41:EGFP+ HSPCs by flow cytometry (FC) at 48 hpf. (D) runx1/cmyb WISH in caspa+/? and caspa?/? embryos at 36 hpf. (E) Phenotypic distribution plot of runx1/cmyb expression scored in embryos from (D) (n ? 46 /condition). (F) CD41:EGFP+ HSPCs in the CHT of embryos from caspa+/? incrosses at 48 hpf. Counts normalized relative to caspa+/+ embryos for each clutch. Significance determined by Tukey?s multiple comparison test. (G) Representative in vivo images of CD41+ HSPCs in the CHT region (brackets) in embryos from (F). (H) pycard knockdown reduces runx1/cmyb expression by WISH (36 hpf; n ? 32 / condition). (I) pycard knockdown decreases the frequency of HSPCs in Tg(flk1:dsRed;cmyb:egfp) embryos by flow cytometry (48 hpf). (J) runx1/cmyb WISH in pycard?/? embryos compared with stage-matched pycard+/+ and pycard+/? embryos (n ? 29/condition). (K) CD41+ HSPCs in the CHT of embryos from pycard+/? incrosses. Counts were normalized relative to pycard+/+ embryos for each clutch. Significance was determined by Fisher?s LSD test. (L) runx1/cmyb expression in the aorta with or without nlrp3l knockdown. (M) Phenotypic distribution plot of embryo cohorts from (L) (n ? 85/condition). (N) nlrp3l knockdown decreases the frequency of HSPCs assayed by flow cytometry at 48 hpf. (O) nlrp3 knockdown reduces the frequency of CD41+ HSPCs at 48 hpf. ?p < 0.05, ??p < 0.01, ???p < 0.001, ????p < 0.0001. Scale bars, 100 ?m. Error bars indicate SD. See also Figure S2.