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Fig. S6
Ccl25b acts as a downstream factor of klf6a in the CHT. Related to Figure 6.
(A) The validation of ccl25b ATG MO. (B) Expression of gata1, mpo and cmyb in control, klf6a morphants and ccl25b morphants at 4.5 dpf (mean ± SD, t-test, **P<0.01, ***P<0.001, n = 3). (C) Expression of cmyb, gata1 and lyz in the CHT of control and ccl25b ATG morphants. (D) The ccl25b splice MO was designed to target the boundary of exon2 and intron 2 (red line marks the target region). The primers were used to amplify the exon region for PCR validation of this MO. (E) Overexpression of ccl25b by injection of hsp-ccl25b-GFP plasmid increased cmyb expression in the CHT. The top right panel showing the GFP expression after heat shock. (F) Expression of cmyb and l-plastin in the CHT of control, ccl25b splice MO-injected embryos and ccl25b splice MO plus ccl25b mRNA-injected embryos at 4 dpf. (G) Confocal imaging of Tg (kdrl:mCherry/runx1:en-GFP) and Tg (kdrl:mCherry/CD41:GFP) showing HSPCs in the CHT of control and ccr7 morphants at 60 hpf and 3 dpf. White arrowheads indicate HSPCs. Scale bar, 50 μm. (H) Expression of cmyb and lyz in control embryos, klf6a morphants and klf6a MO plus ccl25b mRNA injected embryos at 5 dpf. (I) qPCR result of cmyb, gata1, mpo and irf4a in both morphants and klf6a MO plus ccl25b mRNA injected embryos (mean ± SD, t-test, *P<0.05, **P<0.01, ***P<0.001, ns>0.05, n = 3). ns, not significant.
Reprinted from Developmental Cell, 42(4), Xue, Y., Lv, J., Zhang, C., Wang, L., Ma, D., Liu, F., The Vascular Niche Regulates Hematopoietic Stem and Progenitor Cell Lodgment and Expansion via klf6a-ccl25b, 349-362.e4, Copyright (2017) with permission from Elsevier. Full text @ Dev. Cell