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Figure 4 | BMC Developmental Biology

Figure 4

From: A regulatory network of two galectins mediates the earliest steps of avian limb skeletal morphogenesis

Figure 4

Dynamics of the CG-1A and -8 binding reactivity in chicken leg bud mesenchyme. (a) Staining by biotinylated CG-1A for its binding sites in a 3½-day chicken leg section. (b) Staining for CG-1A binding reactivity in 5-day leg sections in the zeugopod (red arrowheads) and in the autopod (blue arrowhead) and a more diffuse spatial localization of CG-8 binding reactivity (c). (d) Staining showing CG-1A binding reactivity in a 6-day leg-bud in the prospective digit primordia (red arrowheads), moderate staining in the interdigital mesenchyme (blue arrowhead) and lack of staining in the interstitial mesenchyme (yellow arrowhead), and (e) more diffuse staining for CG-8 binding reactivity. Images (a-c) have the same scale, as do images (d) and (e), with the scale bars in (a) and (d) representing 1 mm. (f-h) Micromass cultures of leg bud mesenchyme at 15 h of incubation. The culture in (g) shows the spatial pattern of CG-1A binding reactivity using a biotinylated CG-1A probe. The two flanking panels show control cultures. In (f) the biotinylated CG-1A probe was omitted and in (h) the culture was treated with probe that had been pre-incubated overnight with 50 mM lactose. Diameter of the micromass cultures, ~3 mm. (i) Condensation-specific expression of CG-1A-reactive sites in a 2-day leg micromass culture. (j) Addition of 10 μg/ml CG-1A increases extent of its reactivity and expands its localization to precondensation cells, in contrast to the nodular pattern seen in the untreated culture (i). (k) Staining profile of CG-8-reactive sites with biotinylated CG-8 in 2-day leg micromass culture show them to be present both outside and inside condensations. The staining within the condensations is more intense than in the pericondensation regions. Images (i-k) are at the same magnification; scale bar in (i) represents 0.2 mm.

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