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Figure 4. Immunohistochemical (IHC) analysis of Xenopus laevis matrix metalloproteinase 28 (XMMP-28) during Xenopus laevis development. A,D–I,L: Six-micrometer sections of embryos were subjected to IHC to detect XMMP-28. Panels A–C are longitudinal sections of hindlimbs. A: Stage 55 hindlimb. B: Stage 55 hindlimb stained with isotype control antibody to nonexpressed epitope (M2 anti-FLAG, Sigma). C: Stage 55 hindlimb stained with primary XMMP-28 antibody in the presence of 1 μg/ml MMP-28 blocking peptide (Cedarlane). D–F: Stage 53 hindlimb stained for XMMP-28 (D), acetylated tubulin (E) and merged image (F); nuclei were counterstained with DAPI. G–I: Stage 53 brain (G), stage 53 spinal cord (H), stage 53 spinal ganglion (I). J: Whole-mount stage 53 head stained for XMMP-28. K: Whole-mount stage 53 head stained with isotype control antibody. L: Stage 57 hindlimb. Arrows indicate nerves. Scale bars shown in microns. Images representative of two independent experiments. |
