1. Deletion of the Homeobox Gene PRX-2 Affects Fetal but Not Adult Fibroblast Wound Healing Responses 
Philip White, David W. Thomas, Steven Fong, Eric Stelnicki, Fritz Meijlink, Corey Largman, Phil Stephens 
Journal of Investigative Dermatology 
Volume 120, Issue 1, Pages (January 2003)
DOI: /j x
Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Compared to adult fibroblasts fetal fibroblasts demonstrated increased expression of the homeobox gene Prx-2 in monolayer and upregulation in three-dimensional culture. (A) Prx-2 and cytoplasmic actin RNase protection analysis of RNA extracted from fetal or adult wild-type murine fibroblasts (n=3 RNA samples combined) maintained in monolayer culture (MONO), floating collagen lattices (Flt), or attached collagen lattices (Att). (B) Densitometric analysis of Prx-2 RNase protection analysis normalized to actin.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Deletion of the Prx-2 gene reduced fetal but not adult fibroblast proliferation. Fibroblasts isolated from both (A) fetal and (B) adult Prx-2+/+ and Prx-2-/- mice were placed in 96-well microtiter plates at a density of 5×103 cells per well and incubated for 24 or 72 h. Proliferation was assessed by the MTT dye-reduction assay and calculated as the percentage increase in absorbance at day 3 relative to day 1. Values represent the mean±SEM (n≥3).
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Absence of Prx-2 expression had no effect on either fetal or adult cellular attachment to ECM molecules. (A) Fetal and (B) adult Prx-2+/+ and Prx-2-/- murine fibroblasts (1.5×104 cells per well) were added to type I collagen (40 μg per ml), fibronectin (20 μg per ml), or laminin (20 μg per ml) coated wells of a 96-well microtiter plate for 60 min. Following fixation, staining, washing, and extraction the absorbance was measured at 550 nm. Values represent the mean±SEM (n≥4).
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Deletion of the Prx-2 gene increased fetal but not adult fibroblast ECM reorganizational ability independent of cell proliferation. Cultured (A) fetal and (B) adult Prx-2 knockout and wild-type fibroblasts were seeded into type I collagen lattices (starting collagen concentration 1.7 mg per ml) at a cell density of 1×106 cells per 53 mm bacteriologic grade plate. Diametric contraction was measured over a period of 7 d. Values represent the mean±SEM (n≥6). Cultures of (C) fetal and (D) adult Prx-2 knockout and wild-type fibroblasts were seeded into “scaled down” type I collagen lattices at a cell density of 2×104 cells per well of a 24-well plate. At 1, 2, and 7 d five lattices were pooled and subjected to collagenase digestion and viable cell counts were performed. Values represent the mean±SEM (n=3).
Journal of Investigative Dermatology  , DOI: ( /j x)
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6. Figure 5
Absence of Prx-2 expression affects fetal fibroblast MMP levels. Zymography of CM from type I collagen lattices seeded with (A) fetal or (B) adult Prx-2+/+ and Prx-2-/- murine fibroblasts (1×106 cells per lattice). Lanes 1–4, CM from Prx-2 wild-type fibroblasts (days 1, 2, 3, and 7, respectively). Lanes 5–8, CM from Prx-2 knockout fibroblasts at the same time points. Densitometric analysis of (C, E) fetal or (D, F) adult Prx-2+/+ and Prx-2-/- fibroblast relative pro-enzyme and active MMP-2 levels in CM after 7 d in culture in a type I collagen lattice. Values represent the mean±SEM (n=3).
Journal of Investigative Dermatology  , DOI: ( /j x)
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7. Figure 6
Absence of Prx-2 expression had no effect on either fetal or adult fibroblast TIMP levels. Reverse zymography of CM from type I collagen lattices seeded with (A) fetal or (B) adult Prx-2+/+ and Prx-2-/- murine fibroblasts (1×106 cells per lattice). Lanes 1–4, CM from Prx-2 wild-type fibroblasts (days 1, 2, 3, and 7, respectively). Lanes 5–8, CM from Prx-2 knockout fibroblasts at the same time points.
Journal of Investigative Dermatology  , DOI: ( /j x)
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8. Figure 7
Deletion of Prx-2 increases total HA production by fetal, but not adult, fibroblasts but has no effect on HA size. Uronic acid analysis of HA in CM obtained from type I collagen lattices seeded with (A) fetal or (B) adult Prx-2+/+ and Prx-2-/- murine fibroblasts (1×106 cells per lattice) over a period of 7 d. Values represent the mean±SEM (n=3). Gel filtration chromatography elution profiles of CM obtained from type I collagen lattices seeded with (C) fetal or (D) adult Prx-2 knockout and wild-type murine fibroblasts (1×106 cells per lattice) after 7 d. Values represent the mean (n=3; error bars have been excluded for clarity).
Journal of Investigative Dermatology  , DOI: ( /j x)
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9. Figure 8
Absence of Prx-2 has no effect on fibroblast collagen production. The cold hydroxyproline method was utilized to determine the collagen production levels of fetal and adult Prx-2+/+ or Prx-2-/- murine fibroblasts in the presence of serum (10% vol/vol) or TGF-β1 (10 ng per ml). Values represent the mean±SEM (n=3).
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions