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Fig. S5 AM1241 increases HSC number in the AGM in a PGE2-dependent manner, without affecting the vasculature.
(A) In vivo imaging of flk1:dsRed;cmyb:egfp embryos confirmed PGE2 production and signaling are required to mediate the effect of AM1241 on AGM HSCs (n≥10/condition).
(B) In vivo imaging of cd41:gfp embryos confirmed that the AM1241-mediated increase in HSCs in the CHT is independent of PGE2 production and signaling (n≥9/condition).
(C) Western blot analysis showed CNR2-modulation during niche specification altered pERK levels at 24hpf (DMSO: 1 a -actin, n=50 pooled embryos/condition, x 3 replicates).
(D) Western blot analysis confirmed CNR2-modulation altered pAKT protein levels at 24hpf (DMSO: 1-actin, n=50 pooled embryos/condition x 3 replicates).
(E) Flk1:GFP vascular patterning was not impacted by AM1241 exposure (12-30hpf) (n≥20/condition).
(F) Expression of flt4 in the vein was not affected in AM1241-treated embryos (n≥25/condition).
(G) Arterial identity, as indicated by ephrinB2a, was not altered in AM1241-exposed embryos (n≥20/condition).
Scale bars: A,B,G=100µm, E,F=125µm.