IMAGE

Fig. S4

ID
ZDB-IMAGE-150918-9
Source
Figures for Rossi et al., 2015
Image
Figure Caption

Fig. S4 Slc7a7 is expressed in anteriorly located macrophages and is necessary for the establishment of the microglial population. Related to Figure 3 and Figure 4.

(A) Genetic map of the NO067/slc7a7 locus. (B and C) Sequence chromatogram showing the mutation in the slc7a7 coding sequence (ENSDARG00000055226). Sequence analysis identifies an A to T mutation in the fourth splice acceptor of slc7a7 (B) that leads to skipping of the fifth exon (C). Box in (B) indicates the premature stop codon. (D) RT-PCR analysis using primers flanking the exon5 of the slc7a7 cDNA to confirm the efficacy of the morpholino-mediated knockdown. A band shift is present both at 2 and 3 dpf in injected embryos. (E-H) WISH for slc7a7 in a wildtype (E-F) and a pU.1 morphant (G-H) embryo at 2 dpf and 4 dpf (dorsal view). Knockdown of pU.1 leads to lack of slc7a7+ cells in the head (white arrowheads) but not in pancreas and kidney (yellow arrowheads). (I-K) Lateral view of a representative slc7a7::Kaede;pU.1::TagRFP embryo at 2 dpf. Slc7a7+ cells are in green (I, slc7a7::Kaede) and leukocytes are in red (J, pU.1::TagRFP), (K) overlay. Few cells are both pU.1+ and slc7a7+ (yellow arrowheads). White arrowheads indicate pU.1+ cells only. (L-Q) Dorsal view of a representative brain of a 4 dpf slc7a7::Kaede transgenic embryo not photoconverted (L-N) and photoconverted at 44 hpf (O-Q). (L and O) Kaede green, (M and P) Kaede red, (N and Q) overlay. Scale bars for all images: 25 μm.

Acknowledgments
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