Journal:
Article Title: HoxD3 Accelerates Wound Healing in Diabetic Mice
doi:
Figure Lengend Snippet: Expression of HA/HoxD3 in wounds. a: Immunoperoxidase staining for HA in frozen sections from control wounds in db/db mice 7 days after treatment with pellets containing βgal cDNA and stained with a polyclonal antibody against HA. Only background staining (red/brown color) derived from extravasated red blood cells is observed in the tissue. b: Immunoperoxidase staining for HA/HoxD3 in frozen sections from wounds in db/db mice 7 days after treatment with pellets containing an HA epitope-tagged HoxD3 DNA expression plasmid. Tissues were stained with a polyclonal antibody against HA. Positive staining (red/brown color) is observed in numerous areas of granulation tissue (arrows) that has formed at the edge of the wound. c: Higher magnification of control plasmid-treated wounds (a) showing a lack of positive staining in the tissue. d: Higher magnification of HA/HoxD3-treated wounds (b) showing positive cellular staining. e: RT-PCR showing relative levels of HoxD3 mRNA in 7-day wound tissue harvested from wild-type, db/db mice treated with control plasmid or db/db wounds treated with HoxD3. f: Immunoperoxidase staining for HA/HoxD3 in granulation tissue of wounds 5 days after application of HA-HoxD3 plasmids. Positive staining is observed in both fibroblasts and endothelial cells. g: Immunoperoxidase staining for HA/HoxD3 in the looser granulation tissue of 7-day wounds shows an increased number of transfected fibroblasts as compared to 2-day wounds. h: Immunoperoxidase staining for HA/HoxD3 in wounds 7 days after application of HA epitope-tagged HoxD3 DNA. Positive staining is observed in basal keratinocytes (arrow) at the edge of the wound.
Article Snippet: For construction of the HA/HoxD3 expression plasmid, the HoxD3 coding sequence cloned into a CMV-driven expression plasmid (pcDNA3) as previously described, 11 was excised using Kpn / Not I and inserted in frame into the pHM6 epitope expression vector under control of the CMV promoter (Boehringer Mannheim, Indianapolis, IN).
Techniques: Expressing, Immunoperoxidase Staining, Staining, Derivative Assay, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Transfection
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