Figure 2.
Absence of grna leads to decreased myeloid differentiation during embryo development. (A) Representative fluorescence images, and quantification by fluorescence microscopy (A′,A″) of the tails of 48 hpf Mpeg1:eGFP; Lyz:DsRed double transgenic embryos injected with Grna mismatch control, Grna, or Grnb MOs. (B,B′-F,F′) WISH for the myeloid progenitor (pu.1), macrophage (mfap4), neutrophilic (mpx), and microglia (apoeb) markers in grna−/− and grna+/+ control embryos at 48 hpf (B,B″-E,E″) or 5 days post fertilization (dpf) (F-F″). Black arrowheads depict cells expressing the indicated transcripts. (B″,C″,D″,E″,F″) Enumeration of apoeb, pu1+, mfap4+, and mpx+–expressing cells shown in (B-F and B′-F′). Each dot represents the number of positive cells in the photographed area in each embryo. Bars represent mean ± SEM. *P < .05; ***P < .001. (D) Magnification ×10. (G,G′) Maximum projections of the CHT (dotted blue region) of 36 hpf (G) grna+/+ or (G’) grna−/− embryos assayed for TUNEL (red) and 4′,6-diamidino-2-phenylindole (DAPI) (white nuclei). White arrowheads denote apoptotic nuclei. (G″) Enumeration of apoptotic cells in the CHT are quantified in panel G″. Horizontal lines indicate mean ± SEM. ***P < .001. ns, not significant.

Absence of grna leads to decreased myeloid differentiation during embryo development. (A) Representative fluorescence images, and quantification by fluorescence microscopy (A′,A″) of the tails of 48 hpf Mpeg1:eGFP; Lyz:DsRed double transgenic embryos injected with Grna mismatch control, Grna, or Grnb MOs. (B,B′-F,F′) WISH for the myeloid progenitor (pu.1), macrophage (mfap4), neutrophilic (mpx), and microglia (apoeb) markers in grna−/− and grna+/+ control embryos at 48 hpf (B,B″-E,E″) or 5 days post fertilization (dpf) (F-F″). Black arrowheads depict cells expressing the indicated transcripts. (B″,C″,D″,E″,F″) Enumeration of apoeb, pu1+, mfap4+, and mpx+–expressing cells shown in (B-F and B′-F′). Each dot represents the number of positive cells in the photographed area in each embryo. Bars represent mean ± SEM. *P < .05; ***P < .001. (D) Magnification ×10. (G,G′) Maximum projections of the CHT (dotted blue region) of 36 hpf (G) grna+/+ or (G’) grna−/− embryos assayed for TUNEL (red) and 4′,6-diamidino-2-phenylindole (DAPI) (white nuclei). White arrowheads denote apoptotic nuclei. (G″) Enumeration of apoptotic cells in the CHT are quantified in panel G″. Horizontal lines indicate mean ± SEM. ***P < .001. ns, not significant.

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