1. Three-Dimensional Migration of Human Adult Dermal Fibroblasts from Collagen Lattices into Fibrin/Fibronectin Gels Requires Syndecan-4 Proteoglycan 
Fubao Lin, Xiang-Dong Ren, Greiling Doris, Richard A.F. Clark 
Journal of Investigative Dermatology 
Volume 124, Issue 5, Pages (May 2005)
DOI: /j X x
Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Fibroblast three-dimensional migration requires heparan sulfate. Collagen gels containing human adult dermal fibroblasts were treated with or without heparinase I or III. Fibroblast invasive migration from collagen lattices into fibrin gels was visually counted under phase microscopy. The data shown are the mean ± SD of three determinations of one experiment and are representative of three different experiments.
Journal of Investigative Dermatology  , DOI: ( /j X x)
Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Human fibroblast syndecan-4 binds to the Hep II domain of fibronectin (FN). (A) Diagrammatic representation of FN molecular domain structure. (B) 35SO4-labeled syndecan-4 binding to FN functional domains, which had been conjugated to agarose beads. The data shown are mean ± SD of three determinations in this experiment and is representative of three different experiments.
Journal of Investigative Dermatology  , DOI: ( /j X x)
Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Platelet-derived growth factor (PDGF)-induced increase of syndecan-4 protein in fibroblasts. Human adult dermal fibroblasts were treated with PDGF (100 ng per mL) for periods up to 24 h (A) or treated with varying dose of PDGF up to 100 ng per mL for period of 8 h (B). After treatment with 0.2 U per mL heparinase I and III for 2 h at 37°C, whole cell lysates were subjected to SDS-polyacrylamide gel electrophoresis at 20 μg protein/track, transferred to nitrocellulose, and probed with an antibody specific for syndecan-4. The proteins were made visible by the chemiluminescence method. Gels were reprobed with goat antibody specific for human actin to confirm equal loading. The data presented are representative of three separate determinations performed with separate cell lysates.
Journal of Investigative Dermatology  , DOI: ( /j X x)
Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Platelet-derived growth factor (PDGF)-induced increase of syndecan-4 mRNA in fibroblasts. Human adult dermal fibroblasts were treated with or without PDGF (100 ng per mL) in the absence or presence of varying doses of 5, 6 dichlorobenzimidazole 1-β-ribofuranoside for 24 h. The total RNA was extracted with TRIZOL reagent. The levels of syndecan-4 mRNA were determined with Access RT-PCR System using two primers specifically against human syndecan-4 as described in manufacturer's protocols. Equal aliquots of extracted RNA were assayed with primers specific for actin to confirm uniform sampling. The data presented are representative of three separate experiments.
Journal of Investigative Dermatology  , DOI: ( /j X x)
Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
Syndecan-4 antisense oligodeoxyribonucleotides inhibit the transmigration of fibroblasts. Human adult dermal fibroblasts were treated with or without syndecan-4 (AS) or its scrambled antisense (SAS) oligodeoxyribonucleotides for 48 h in the presence or absence of 100 ng per mL platelet-derived growth factor (PDGF). At the conclusion of incubation cells were dissolved in affinity bead binding buffer. After centrifugation, the supernatants were incubated with affinity agarose beads to which syndecan antibodies had been conjugated. Bound syndecans were subjected to SDS-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with goat antibodies specific for human syndecan-4 (A) or human syndecan-2 (B). Gels were reprobed with goat antibodies specific for human actin to confirm equal loading. The proteins were made visible by the chemiluminescence method. The data presented are representative of three separate determinations performed with separate cell lysates. (C) Cells were treated with or without AS or SAS oligodeoxyribonucleotides for 48 h were subjected to 100 ng per mL PDGF in the transmigration assay (see Materials and methods). All data presented are means ± SD from three determinations of one experiment and representative of three experiments performed with cells from different antisense treatments.
Journal of Investigative Dermatology  , DOI: ( /j X x)
Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

7. Figure 6
Syndecan-2 antisense (AS) oligodeoxyribonucleotides fails to inhibit transmigration of fibroblasts. (A) Human adult dermal fibroblasts were treated with or without AS oligodeoxyribonucleotides or scrambled antisense (SAS) oligodeoxyribonucleotides for 48 h in the presence or absence of 100 ng per mL platelet-derived growth factor (PDGF). At the conclusion of incubation cells were dissolved in affinity bead binding buffer. After centrifugation, the supernatants were incubated with affinity agarose beads to which syndecan-2 antibodies had been conjugated. Bound syndecans were subjected to SDS-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with goat antibodies specific for human syndecan-2. Gels were reprobed with goat antibodies specific for human actin to confirm equal loading. Syndecan-2 and actin were made visible by the chemiluminescence method. The data presented are representative of three separate determinations performed with separate cell lysates. (B) Cells were treated with or without AS or SAS oligodeoxyribonucleotides for 48 h were subjected to 100 ng per mL PDGF in the transmigration assay (see Materials and methods). All data presented are means ± SD from three determinations of one experiment and representative of three experiments performed with cells from different antisense treatments.
Journal of Investigative Dermatology  , DOI: ( /j X x)
Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions