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Fig. 2 | Journal of Translational Medicine

Fig. 2

From: Matrix- and plasma-derived peptides promote tissue-specific injury responses and wound healing in diabetic swine

Fig. 2

Cytological and biochemical characteristics of FITC-Comb1 interactions. a FITC-conjugated Comb1 binds to blood vessels and keratinocytes. Fresh frozen sections from diabetic porcine wounds were blocked with 5 % normal goat serum in PBS, then incubated in block solution alone or containing 100 nM FITC-Comb1, followed by immunofluorescence microscopy using anti-FITC (1:200, Life Technologies). Image analysis reveals that FITC-Comb1 binds to wound-associated keratinocytes, stromal, and vascular cells, as well as keratinocytes distal to the cutaneous defect. Image of trichrome-stained tissue provided for location context. Scale bar denotes 500 μm in all panels. b FITC-conjugated Comb1 ligates with numerous proteins expressed in HMVEC, HaCaT, and HFF. Whole cell lysates were prepared from cells denuded from post-confluent monolayers, representing the control/resting state of the cells, and actively recovering cells collected 24 h after injury, then separated by SDS-PAGE. Gels were transferred to nitrocellulose membranes, overlaid with 200 nM FITC-Comb1, and cross-linked to putative receptors and interactors using 2 mM EDC and 2 mM NHS. Interactors were detected using by western blotting with an anti-FITC antibody

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