SEARCH

SEARCH BY CITATION

Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
DVDY_22678_sm_suppfig1.tif11025KSupporting Figure 1 Analysis of reporter gene expression in other tissues than the pectoral fins: the mPrx1(cFos):EGFP lines. A–E: mPrx1(cFos):EGFP line#1. Apart from the expression in the developing pectoral fins from 27 hours postfertilization (hpf)onwards (marked here by arrowheads in C–E), enhanced green fluorescent protein (EGFP) signal is observed in the following territories: (i) In the pronephric duct from 18 hpf (arrows in A–E). (ii) From 30 hpf, EGFP labels some mesenchymal cells in the median fin fold (double arrowheads in C–E). (iii) Some cells of the spinal cord at the level of the pectoral fin from 27 hpf onward (asterisks in C–E and Fig. 2 A, B). (iv) From 48 hpf, expression of EGFP is observed in anterior brain and retina. F–J: mPrx1(cFos):EGFP line#2. Apart from the expression in the developing pectoral fins from 27 hpf onward (marked here by arrowheads in H–J), EGFP signal is observed in the following areas: (i) At 48 hpf, EGFP labels some mesenchymal cells in the median fin fold (double arrowheads in I) and (ii) in mesenchymal cells in the dorsal aspect of the head at 48 and 72 hpf (arrows in I, J). K–O: mPrx1(cFos):EGFP line#5. Apart from the expression in the developing pectoral fins from 24 hpf onward (marked by arrowheads in L–O), EGFP signal is also observed in a dorsal cranial bone of the 48 and 72 hpf embryos marked by arrows in N and O. Specific sites of reporter gene expression (other than the limb) in each of the transgenic lines generated may represent positional effects due to different integration sites of the transgene. Reporter gene expression is also detected in areas of the ventral head mesenchyme in all the transgenic lines created which is a characteristic of the mPrx1 limb element (Logan et al., 2002). Nonspecific expression in the dorsal-most aspect of the yolk extension is observed in all three transgenic lines (A–O).
DVDY_22678_sm_suppfig2.tif9630KSupporting Figure 2 Analysis of reporter gene expression in other tissues than the pectoral fins: the zfprrx1aCNE:EGFP lines. A–E: zfprrx1a(cFos):EGFP line#2. Apart from the expression in the developing pectoral fins from 27 hours postfertilization (hpf) onward (marked by arrowheads in C–E), enhanced green fluorescent protein (EGFP) signal is observed in (i) part of the somites of the 18 hpf embryo (so, in A) and in (ii) the otic vesicle and anterior lateral line from 24 hpf onward (arrows in B–E and Fig. 5 A–C). F–J: zfprrx1a(cFos):EGFP line#6. Apart from the expression in the developing pectoral fins from 27 hpf onward (marked here by arrowheads in H–J), EGFP signal is also observed in the anterior brain and retina from 24 hpf onward (arrows in G–J). The specific sites of reporter gene expression in each of the transgenic lines may represent positional effects due to different integration sites of the transgene. Reporter gene expression is also detected in common areas in the two transgenic lines, which are some mesenchymal cells in the median fin fold and scattered cells surrounding the somatic tissue (mff and double arrowheads(Logan et al., 2002). Nonspecific expression in the dorsal-most aspect of the yolk extension is observed in these transgenic lines (A–J).
DVDY_22678_sm_suppfig3.tif9239KSupporting Figure 3 Analysis of reporter gene expression in other tissues than the pectoral fins: the zfprrx1b1CNE:EGFP lines. A–E: zfprrx1b1(cFos):EGFP line#5. Expression in this transgenic line is very pectoral fin-specific until at least 30 hours postfertilization (hpf; Fig. 6 and C here). F–J: zfprrx1b1(cFos):EGFP line#8. Apart from the expression in the developing pectoral fins from 27 hpf onward (marked here by arrowheads in H–J), enhanced green fluorescent protein (EGFP) signal is also observed in some cells of the spinal cord from 18 to 48 hpf (arrows in F–I). Because this expression site is not observed in the other transgenic line generated by injection of the same DNA construct (zfprrx1b1(cFos):EGFP line#5), it is most probably due to a positional effect. Similar to the expression observed in the zfprrx1aCNE:EGFP lines EGFP signal is also observed in scattered cells surrounding the somatic muscles at 48 and 72 hpf (double arrows in D, E and I, J), suggesting these represent true expression sites driven by the orthologous enhancers zfprrx1a and zfprrx1b1 CNEs. Again, nonspecific expression in the yolk extension is observed (A–J).
DVDY_22678_sm_suppfig4.tif9357KSupporting Figure 4 Analysis of reporter gene expression in other tissues than the pectoral fins: the zfprrx1b2CNE:EGFP lines. A–E: zfprrx1b2(cFos):EGFP line#11. Apart from the expression in the developing pectoral fins from 24 hours postfertilization (hpf)onward (marked by arrowheads in B–E), detectable enhanced green fluorescent protein (EGFP) signal is observed in (i) the notochord from 18 to 30 hpf (nc, A–C) and in scattered cells surrounding the somatic muscles at 72 hpf (double arrows in E). F–J: zfprrx1b2(cFos):EGFP line#13. Apart from the expression in the developing pectoral fins from 24 hpf onward (marked here by arrowheads in G–J), EGFP signal is also observed in the following territories: (i) in the posterior-most somites from 18 to 30 hpf (so, F–H), (ii) in the hatching gland from 18 hpf onward (hg, F–I), (iii) in the otic vesicle from 24 hpf onward (ov, G–I), (iv) in the spinal cord from 30 hpf (arrows in H–J) and (v) in the anterior brain and retina as well as mesenchymal cells of the medial fin fold of the 48 and 72 hpf embryos (mff, I, J). The specific sites of reporter gene expression in the transgenic lines reported here represent positional effects due to different integration sites of the transgene. Nonspecific expression in the dorsal-most aspect of the yolk extension is also observed in these transgenic lines (A–J). All the images shown in Supp. Figs. 1S–S4 have been enhanced (as a whole) to make EGFP expression in other sites than the fins clearly observable.
DVDY_22678_sm_suppfig5.tif3633KSupporting Figure 5 Original images with the same settings as those used for Figure 2. A: zfprrx1a CNE line#2 27 hpf embryo. B: zfprrx1a CNE line#2 30 hpf embryo. C: zfprrx1a CNE line#6 27 hpf embryo. D: zfprrx1a CNE line#6 30 hpf embryo. E: zfprrx1b1 CNE line#5 27 hpf embryo. F: zfprrx1b2 CNE line#11 24 hpf embryo. Expression in the pectoral fins is marked by arrowheads.

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.