Journal: Cancer Medicine

Article Title: Sam68 is required for the growth and survival of nonmelanoma skin cancer

doi: 10.1002/cam4.2513

Figure Lengend Snippet: Sam68 protein level is elevated in human and mouse skin cancers. A, Immunofluorescence micrographs of Sam68, with nuclei counterstained by DAPI, on skin tissue sections from healthy controls or nonmelanoma skin cancer (NMSC) patients consisting of both squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) cases. T, tumor; derm, dermis; epi, epidermis. Scale bars, 100 μm. Right , the fluorescence intensity (FI) of Sam68 on the indicated skin tissue sections was quantified. B, Hematoxylin and eosin staining of human healthy control skin, SCC, and BCC tissues. Scale bars, 500 μm. C, Immunofluorescence micrographs of Sam68, with nuclei counterstained by DAPI, on normal and tumor skin tissue sections from wild‐type (WT) and tumor‐laden Gli2 tg/+ mice, respectively. T, tumor; derm, dermis; epi, epidermis. Scale bars, 50 μm. D, Whole cell lysates derived from adjacent normal (N) and tumor (T) skin tissue from the same tumor‐laden Gli2 tg/+ mice at postnatal day 60 were immunoblotted (IB) for Sam68, with Histone H3 as a loading control. Bottom , normalized Sam68 level in the N and T skin tissues in Gli2 tg/+ mice (n = 3) was quantified. E, Skin tissue was collected from wild‐type mice or Gli2 tg/+ mice at the indicated postnatal days and whole cell lysates were derived (each sample indicates individual mouse) and IB for Sam68, with β‐actin as a loading control. The normalized Sam68 level in the skin of indicated mice was quantified on the bottom. Protein expression levels were measured from at least three mice per genotype/time point. AU, arbitrary unit. Data information: In (A, D‐E), data are presented as mean ± SEM. * P < .05; ** P < .01; and *** P < .001 (Student's t test)

Article Snippet: Human skin cancer tissue arrays were purchased from US Biomax Inc, where all human tissues have been collected under HIPAA‐approved protocols with donors being informed in full and with their consent.

Techniques: Immunofluorescence, Fluorescence, Staining, Derivative Assay, Expressing

Journal: Cancer Medicine

Article Title: Sam68 is required for the growth and survival of nonmelanoma skin cancer

doi: 10.1002/cam4.2513

Figure Lengend Snippet: Sam68 plays a critical role for mouse development of skin cancer. A, Representative macrographs of P70 ears from the mice with indicated genders and genotypes. Arrows point to blood vessels. Scale bars, 1 cm. B, The tumor lesion onset kinetics in Gli2 tg/+ ; Sam68 ± and Gli2 tg/+ ; Sam68 −/− mice. ** P < .02 by Gehan‐Breslow‐Wilcoxon test. C, Hematoxylin and eosin staining of P70 ear tissue sections derived from mice with indicated genotypes. Scale bar, 1 mm. D, Immunofluorescence micrographs of PECAM‐1, with nuclei counterstained by DAPI, on P70 ear tissue sections from Gli2 tg/+ ; Sam68 ± and Gli2 tg/+ ; Sam68 −/− mice. T, tumor; derm, dermis; epi, epidermis. Scale bars, 100 μm. Data information: In (B), ** P < .02 (Gehan‐Breslow‐Wilcoxon test)

Article Snippet: Human skin cancer tissue arrays were purchased from US Biomax Inc, where all human tissues have been collected under HIPAA‐approved protocols with donors being informed in full and with their consent.

Techniques: Staining, Derivative Assay, Immunofluorescence

Journal: Molecular Carcinogenesis

Article Title: Loss of Desmocollin 3 in Skin Tumor Development and Progression

doi: 10.1002/mc.20818

Figure Lengend Snippet: Loss of DSC3 expression in control mouse skin tumors. (A-G) Immunofluorescence microscopy staining of sections through a well-differentiated control (Dsc3fl/+; K-RasG12D; K5.Cre*PR1) tumor. The tumor sections were stained with the antibodies indicated on the left. (A) Note that DSC3 is present at cell-cell borders in the hyperplastic epithelium (HE) overlaying the tumor tissue (T). The tumor has lost DSC3 staining at the cell-cell borders. The remaining weak signal in the cytoplasm most likely represents background staining. (B-E) Other desmosomal markers, such as desmogleins 3 (DSG3), plakophilin 3 (PKP3; a cytoplasmic binding partner for DSC3), desmoplakin (DSP) and plakoglobin (JUP) show normal expression patterns in the tumor tissues. Note that desmosomal proteins normally show a cell type specific expression patterns. For example, DSG3 expression is restricted to the basal and first suprabasal cell layers in normal epidermis and in the tumor tissue (arrow in (B)). (F) The tumor progression marker KRT13 is expressed in tumor tissue that has lost DSC3 expression (compare consecutive tissue sections in (A) and (F)]. (G) Staining with normal guinea pig IgG (negative control staining for section (A)). Counterstaining with an antibody against α6 integrin (ITGA6) demarcates the basement membrane (red). Bar, 50μm. (H) Histological section through the tumor shown in (A) – (G). (I) Histological section through the tumor shown in (J-K). In situ hybridization with Dsc3 (J) anti-sense and (K) sense probes on control tumor sections. The basement membrane zone of the overlaying HE is marked with a dashed line. Note that the hyperplastic epithelium (HE) is positive for Dsc3 mRNA while part of the tumor tissue (T) is negative (arrow in J), indicating loss of Dsc3 gene expression. The sense probe did not hybridize to the tissue section, demonstrating specificity of the ani-sense hybridization for Dsc3 mRNA. Bar, 50μm. (L) Quantification of the immunofluorescence signals obtained with the antibodies shown in A-E. As outlined in MATERIAL AND METHODS, the fluorescence signals in hyperplastic and tumor areas of the same tumor were determine and compared to each other. Expression levels of DSC3 were significantly reduced in the tumor tissue. Note that due to the diffuse background staining in the tumor portion of A, we are likely to overestimate residual DSC3 staining. Note the small but statistically significant reduction in PKP3 expression in the tumor tissue when compared to the hyperplastic epithelium.

Article Snippet: Human Tissue Arrays The human skin cancer tissue array was purchased from US Biomax (Rockville, MD.

Techniques: Expressing, Immunofluorescence, Microscopy, Staining, Binding Assay, Marker, Negative Control, In Situ Hybridization, Hybridization, Fluorescence

Journal: Molecular Carcinogenesis

Article Title: Loss of Desmocollin 3 in Skin Tumor Development and Progression

doi: 10.1002/mc.20818

Figure Lengend Snippet: DSC3 expression in human skin SCCs. A human skin tumor tissue array (see MATERIAL AND METHODS) was co-stained with antibodies against keratin 5 (KRT5, red) and DSC3 (green). (A) As expected, both antibodies stained normal human skin. (B-D) Skin SCC sections showed a range of DSC3 expression patterns including (B) strong homogeneous expression (Grade 1 SCC), (C) patchy expression pattern (DSC3-positive and DSC3-negative tumor areas within the same section; Grade 2 SCC), (D) a complete loss of DSC3 expression in KRT5-positive cells (Grade 2 SCC). Note that the DSC3-stained sections were photographed with the same exposure time. Tumor tissues were classified by the provider of the array (see MATERIAL AND METHODS). Bar, 50μm. (E) Percentage of tissue sections showing either homogeneous or reduced/absent staining for DSC3 in KRT5-positive cells.

Article Snippet: Human Tissue Arrays The human skin cancer tissue array was purchased from US Biomax (Rockville, MD.

Techniques: Expressing, Staining