18 research outputs found
Spatial relationship of Wnt5a, Fzd3, and Fzd5 in squamous cell carcinoma.
<p>Serial sections of three paraffin embedded SCC tumor samples (top, middle, bottom row, respectively) were stained for Wnt5a, Fzd5, Fzd3, respectively, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031827#s2" target="_blank">Methods</a>, and shown at 200× magnification. Red asterisk denotes artificial nuclear staining possibly due to antigen retrieval conditions. Red arrows denote boundaries of tumors, pointing toward stroma.</p
Absence of inflammatory changes induced by PPAR β/δ antagonists in skin after topical application.
<p>(a) C57Bl/6j wild type mice were treated with ointments containing GSK0660 or compound 3 h applied twice daily to shaved dorsal skin for one week. Mice were sacrificed 1 h after the last ointment application and skin tissue processed for H&E based histology and mass spectrometry, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037097#s4" target="_blank">Methods</a>. Data shown represent average ± s.d. of n = 3 mice per data point (left) treated with GSK (blue columns) or compound 3 h (red). Representative histology sections of all treated mice are shown on right. The inset in the middle panel shows a section of GSK0660-treated epidermis showing apoptotic looking cells (marked by red arrow head). Horizontal bar represents 5 µm. (b) Representative H&E sections of C57Bl/6j wild type mice treated for one week with either GSK0660 (top) or GSK3787 (bottom). Red arrow-heads denoting apoptotic looking cells.</p
Immunohistochemical detection of β-catenin in BCC (b, e), and moderately differentiated SCC (c, f) samples at the ProteinAtlas repository (see main text).
<p>Samples were stained either with an antibody specific for activated non-phosphorylated β-catenin (top) or pan-β-catenin (bottom). Images in (a) and (d) show the β-catenin distribution observed with the respective antibody. Note that strong nuclear β-catenin is confined to the granular layer of the epidermis.</p
Expression of Wnt5a, Fzd3, and Fzd5 in non-melanoma skin cancer.<sup>1</sup>
1<p>Immunohistochemistry of formaldehyde-fixed paraffin-embedded SCC (n = 12) and BCC (n = 9) samples was carried out as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031827#s2" target="_blank">Methods</a>.</p>2<p>Expression level was scored as “moderate” when staining intensity was comparable, as “strong” when staining was stronger, and as “low” when staining was weaker than that of epidermis present in the same section, respectively.</p
Wnt5a inhibits keratinoctye migration when present in homogenous concentration, but acts as chemoattractant when present as gradient.
<p><b>A.</b> Expression of endogenous and recombinant Wnt5a in whole cell lysates of stably transfected Wnt5a-overexpressing HaCat or control (HaCat-pcDNA) cells verified by western blot. <b>B.</b> Non-Wnt5a overexpressing HaCat-pcDNA cells were seeded in the upper chamber of a Transwell in 0.1% BSA DMEM in the absence or presence of recombinant Wnt5a at 1 µg/ml, as indicated in the figure. The lower chamber was filled with 600 µl DMEM containing 5% FCS as chemoattractant. Results are expressed as percentage of migrating cells when HaCat-pcDNA were seeded in 0.1% BSA DMEM only. The results shown represent mean ± s.d. of two independent experiment, each performed in triplicate, *p≤0.05. <b>C.</b> Comparison of Wnt5a-overexpressing and pcDNA control cell migration. Cells suspended in 0.1% BSA DMEM were seeded in the upper chamber. The lower chamber were filled with 600 µl DMEM containing 5% FCS as chemoattractant. Migration was assessed at 18 h using a colorimetric assay. Results are expressed as percentage of HaCat-pcDNA migrating cells. Results shown represent mean ± s.d. of n = 4 independent experiment, each performed in triplicate, *** p≤0.001. <b>D.</b> Scratch wound assay performed on mitomycin-C treated cells. During migration, HaCat-pcDNA (a, b, c), or Wnt5a-overexpressing cells (d, e, f) were maintained in DMEM containing 10% FCS. Pictures were taken just after the scratch was made (0 hrs) (a and d), as well as 18 h (b and e) and 24 h later (c and f). <b>E.</b> Migration of HaCat-pcDNA control cells in the presence of a Wnt5a concentration gradient. Wnt5a-overexpressing or pcDNA HaCat cells were seeded in the bottom wells of Transwell plates. Immediately before adding the inserts containing HaCat-pcDNA cells in the upper chamber, the media in the bottom wells was replaced to remove pre-secreted Wnt5a. Migration was assessed at 18 h. Results are expressed as percentage of HaCat-pcDNA migrating cells. Results shown represent mean ± s.d. of n = 3 independent experiments, each performed in triplicate, *** p≤0.001.</p
Spatial relationship of Wnt5a, Fzd3, and Fzd5 localization in basal cell carcinoma.
<p>Immunohistochemistry of serially cut samples stained for Wnt5a, Fzd5, or Fzd3 as indicated, magnification: 100× (top row), 200× (middle, bottom rows).</p
Expression of Wnt – ligands in cutaneous SCC.<sup>1</sup>
1<p>Data shown (extracted from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031827#pone.0031827-Hudson1" target="_blank">[18]</a>) represent n = 10 paired samples of moderately differentiated cutaneous invasive SCC from immunocompetent patients. BOLD print: fluorescence intensity (arbitrary units) of >1000 in either control or tumor.</p>2<p>Expression levels are the inverse log2-transform of raw fluorescence intensity data provided by <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031827#pone.0031827-Hudson1" target="_blank">[18]</a>. Where multiple probes per gene were present on the array, the probe yielding the highest fluorescence value is shown. For none of the genes listed in the table was there an inverse or significant fold-change for alternative probes.</p>3<p>Fold-changes between SCC and control samples with a p-value <0.02 are shown.</p>4<p>Classified according to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031827#pone.0031827-Yuzugullu1" target="_blank">[27]</a>.</p
Localization of Fzd5 in non-melanoma skin cancer.
<p>Immunohistochemistry of SCC (a–d), or BCC (e–h), in each case showing an example of tumors exhibiting high (SCC: a,c; BCC: e,g) or low (SCC: b,d; BCC: f,h) Fzd5 expression. Staining intensities in the tumors can be directly compared to staining intensities of Fzd5 in the granular layer of the epidermis (black arrowheads). Panels a,b,e,f are shown at 40× and c,d, g,h at 200× magnification. The inset on the lower right of (c) shows a tumor-associated fibroblast. Red arrows denote blood vessels.</p
Prevention of epidermal hyperplasia by transdermal application of selective PPAR β/δ antagonists.
<p>Both the PPAR β/δ agonist GW501516 (GW) and the antagonists GSK0660 (GSK) or compound 3 h were applied topically to the skin, as described in the text. Left: representative H&E-stained paraffin-sections of dorsal skin from PPAR β/δ transgenic mice after treatment with ointments containing the indicated drugs for twenty days, as detailed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037097#s4" target="_blank">Methods</a>. Horizontal bar represents 5 µm. Right: quantification of H&E-based epidermal thickness observed in n = 4 mice per group, performed as detailed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037097#s4" target="_blank">Methods</a>. * p<0.05 in a two-sided independent t-test.</p
Low systemic absorption of topically applied PPAR β/δ antagonists.
<p><b>A</b>. Peak blood concentrations of PPAR β/δ agonist GW501516, and antagonists GSK0660 and compound 3 h, respectively, at 1 h after topical application to skin. Left: Amount of drugs detected in systemic circulation, expressed as fraction of total amount applied, was calculated as detailed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037097#s4" target="_blank">methods</a>. Right: Drug concentration expressed as molar concentration. <b>B</b>. GSK0660 concentration in blood (left) and total amount of circulating drug as fraction of amount applied (right, calculated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037097#s4" target="_blank">Methods</a>) at the indicated time points after drug application. The horizontal dashed line represents the reported IC50 for GSK0660 acting on PPAR β/δ reported previously (300 nmol/L). Data shown represent average ± s.d. of n = 3 mice per data point.</p