Journal: Frontiers in Cell and Developmental Biology

Article Title: MicroRNA-124-3p Plays a Crucial Role in Cleft Palate Induced by Retinoic Acid

doi: 10.3389/fcell.2021.621045

Figure Lengend Snippet: Inhibition of miR-124-3p expression rescues at RA-induced CP in mice. (A) Schematic experimental design. (B) Gross picture of E18.5 C57BL/6J mice treated with control vehicle, at RA, or at RA + miR-124-3p inhibitor. Bars, 1 mm. (C) H&E staining of the face of E18.5 C57BL/6J mice treated with control vehicle, at RA, or at RA + miR-124-3p inhibitor. Bars, 400 μm. (D) Ki-67 staining (green) in the developing palate of E13.5 C57BL/6J mice treated with/without a miR-124-3p inhibitor under at RA condition. Nuclei were counterstained with DAPI (blue). Representative images from two independent experiments are shown. Graph shows the quantification of Ki-67 positive cells. One-way ANOVA with Tukey’s honest significant difference test. ** p < 0.01 vs. control. ## p < 0.01 vs. at RA. Bars, 50 μm. (E) Quantitative RT-PCR for the indicated genes after treatment with/without a miR-124-3p inhibitor under at RA condition in the palatal shelves of E14.5 C57BL/6J mice. Two-way ANOVA with Tukey’s honest significant difference test ( n = 3). *Adjusted p < 0.00313 (16 genes), **adjusted p < 0.000625 (16 genes), and ***adjusted p < 0.0000625 (16 genes) vs. control. # Adjusted p < 0.00313 (16 genes), ## adjusted p < 0.000625 (16 genes), and ### adjusted p < 0.0000625 (16 genes) vs. at RA. (F) AZAN staining (upper panels) and immunohistochemical staining for VCAN (middle panels; brown) and CDC42 (bottom panels; brown) in E13.5 C57BL/6J mice treated with control vehicle, at RA, or at RA + miR-124-3p inhibitor. Bars, 25 μm.

Article Snippet: After antigen retrieval treatment with citrate buffer (pH 6.0) and blocking of endogenous peroxidase with 0.3% hydrogen peroxide in methanol, the sections were incubated with anti-cytokeratin 14 mouse monoclonal antibody (Abcam, ab7800, 1:200 dilution), anti-Ki-67 rabbit monoclonal antibody (Abcam, ab16667, 1:200 dilution), anti-VCAN rabbit polyclonal antibody (Novus Biologicals, Centennial, CO, United States, NBP1-85432, 1:200 dilution), or anti-CDC42 rabbit polyclonal antibody (Proteintech, Rosemont, IL, United States, 10155-1-AP, 1:50 dilution) at 4°C overnight.

Techniques: Inhibition, Expressing, Control, Staining, Quantitative RT-PCR, Immunohistochemical staining

Journal: Glycoconjugate Journal

Article Title: Molecular expression of glycosaminoglycans modifies the plasticity of biphasic mesothelioma in favor of tumor progression

doi: 10.1007/s10719-025-10192-z

Figure Lengend Snippet: Histological illustration of epithelioid malignant pleural mesothelioma (E-MPM) and biphasic MPM (B-MPM) tissues. Examples of E-MPM and B-MPM subtypes as defined by heparan sulfate, chondroitin sulfate, versican, byglican, and perlecan expression. Two types of regions are represented based on areas with high expression levels of glycosaminoglycan GAG markers. E-MPM and B-MPM corresponded to areas of pure E- and B- components, respectively. As is shown in Table , low expression levels of chondroitin sulfate and versican were observed in epithelioid cells from E-MPM tumors. Heparan sulfate and byglican were positively expressed by fusiform cells in B-MPM tumors, contrasting with weak expression in E-MPM tumors and E- areas of B-MPM ones. Perlecan expression indicated high levels of GAG markers in both E-MPM and B-MPM tumors. Magnification: 400×. Scale: 50 μm. *** P < 0.001, **** P < 0.0001. The red square in the boxplots represents the mean value of the data

Article Snippet: The slides were stained with immunoperoxidase and antibodies against heparan sulfate (1:100; AMSBIO LLC, #370255-1, clone F58-10E4), chondroitin sulfate (1:200; Sigma-Aldrich, #C8035, clone CS-56), versican (1:100; Santa Cruz Biotechnology Inc, sc-25831, polyclonal), biglycan (1:100; Santa Cruz Biotechnology Inc., sc-33788, polyclonal), perlecan (1:100; Santa Cruz Biotechnology Inc., sc-25848, polyclonal), and elastin (1:100; Santa Cruz Biotechnology Inc., sc-58756, clone BA-4).

Techniques: Expressing

Journal: Glycoconjugate Journal

Article Title: Molecular expression of glycosaminoglycans modifies the plasticity of biphasic mesothelioma in favor of tumor progression

doi: 10.1007/s10719-025-10192-z

Figure Lengend Snippet: Spearman correlation matrix of glycosaminoglycans, proteoglycans, and extracellular matrix (ECM) proteins in malignant pleural mesothelioma tissue samples. The illustration shows the Spearman correlation coefficients (ρ) - represented by the circles, that when larger, the values closer to|1| (i.e., a stronger correlation). Positive correlations are shown in blue (stronger intensity indicates higher correlation), while negative correlations are shown in orange. Statistically significant moderate positive correlations were observed between HS and CS, HS and versican, HS and biglycan, HS and perlecan, versican and biglycan, versican and elastin, and biglycan and elastin. A moderate inverse correlation was noted between perlecan and collagen V. * P < 0.05; ** P < 0.01

Article Snippet: The slides were stained with immunoperoxidase and antibodies against heparan sulfate (1:100; AMSBIO LLC, #370255-1, clone F58-10E4), chondroitin sulfate (1:200; Sigma-Aldrich, #C8035, clone CS-56), versican (1:100; Santa Cruz Biotechnology Inc, sc-25831, polyclonal), biglycan (1:100; Santa Cruz Biotechnology Inc., sc-33788, polyclonal), perlecan (1:100; Santa Cruz Biotechnology Inc., sc-25848, polyclonal), and elastin (1:100; Santa Cruz Biotechnology Inc., sc-58756, clone BA-4).

Techniques:

Journal: Glycoconjugate Journal

Article Title: Molecular expression of glycosaminoglycans modifies the plasticity of biphasic mesothelioma in favor of tumor progression

doi: 10.1007/s10719-025-10192-z

Figure Lengend Snippet: Kaplan–Meier overall survival analysis according to extracellular matrix component expression in malignant pleural mesothelioma patients. Kaplan–Meier survival curves comparing overall survival (OS) between patients stratified by high and low immunohistochemical expression levels of ( A ) heparan sulfate, ( B ) chondroitin sulfate, ( C ) versican, ( D ) biglycan, and ( E ) perlecan. Patients were dichotomized based on median expression cutoffs for each marker. No statistically significant differences in OS were observed between the groups using the log-rank test

Article Snippet: The slides were stained with immunoperoxidase and antibodies against heparan sulfate (1:100; AMSBIO LLC, #370255-1, clone F58-10E4), chondroitin sulfate (1:200; Sigma-Aldrich, #C8035, clone CS-56), versican (1:100; Santa Cruz Biotechnology Inc, sc-25831, polyclonal), biglycan (1:100; Santa Cruz Biotechnology Inc., sc-33788, polyclonal), perlecan (1:100; Santa Cruz Biotechnology Inc., sc-25848, polyclonal), and elastin (1:100; Santa Cruz Biotechnology Inc., sc-58756, clone BA-4).

Techniques: Expressing, Immunohistochemical staining, Marker

Journal: Matrix biology plus

Article Title: Temporal expression and spatial distribution of the proteoglycan versican during cardiac fibrosis development.

doi: 10.1016/j.mbplus.2023.100135

Figure Lengend Snippet: Fig. 2. Versican expression is increased and precedes collagen expression in the early phase of pressure overload. (A) mRNA expression of versican (Vcan, measured using primer pairs targeting all isoforms of Vcan), (B) collagen I alpha 2 (Col1a2), and (C) collagen III alpha 1 (Col3a1) in the left ventricles of mouse hearts at day 1, 3, 7, 14, 28 and 56 after aortic banding (AB, n = 8–11) or sham operations (Control, n = 9–10). The absolute quantity of gene expression (copies/μl measured by ddPCR) relative to respective Control at day 1 is presented. Data represent mean ± SD. Repeated measures two-way ANOVA with Bonferroni’s multiple comparisons test was used for statistical analysis. P values < 0.05 were considered statistically significant. *P < 0.05 Control vs. AB, †P < 0.05 AB vs. day 1 post AB, ‡P < 0.05 AB vs. day 3 post AB.

Article Snippet: The sections were blocked in Protein Block (Serum-Free, Agilent Dako, #X090930-2) for 10 min and incubated overnight with primary antibodies rabbit anti-mouse versican (GAG-β) antibody (Merck Sigma, Germany, #AB1033, 1:1000), rat anti-human versican antibody (R&D systems, MN, #MAB3054, 1:100), and rabbit anti-mouse/human versican V0, V1 neoepitope antibody (Invitrogen, #PA1-1748A, 1:1000) at 4◦C.

Techniques: Expressing, Control, Gene Expression

Journal: Matrix biology plus

Article Title: Temporal expression and spatial distribution of the proteoglycan versican during cardiac fibrosis development.

doi: 10.1016/j.mbplus.2023.100135

Figure Lengend Snippet: Fig. 3. V0 and V1 are the main isoforms expressed during fibrosis development during pressure overload. (A-D) mRNA expression of versican isoforms V0, V1, V2 and V3 from the left ventricle of mouse hearts at day 1, 3, 7, 14, 28 and 56 after aortic banding (AB, n = 8–11) or sham operations (Control, n = 9–10). (E-G) Representative western blot image and quantification (normalized to total protein stain, relative to day 1 post AB) of the V0 and V1 isoform from the left ventricles of mice at day 1, 3, 7, 14, 28 and 56 after aortic banding (AB, n = 8) or sham operations (Control, n = 5–8). All samples were treated with chondroitinase ABC enzyme to remove glycosaminoglycan chains, except for the sample on the right side of the image. Pooled extracellular matrix fractions from left ventricular tissue of aortic banded mice obtained after 8 to 10 weeks were used as positive controls. Control samples (sham operation) displayed very low or no amount of versican. The absolute quantity of mRNA expression (copies/μl measured by ddPCR) relative to mRNA expression measured by primer pairs targeting all isoforms of Vcan is presented. Data represent mean ± SD. Repeated measures two-way ANOVA (A-D) or one-way ANOVA (F) with Bonferroni’s multiple comparisons test, and Kruskal- Wallis (G) with Dunn’s post-hoc test were used for statistical analysis. P values < 0.05 were considered statistically significant. *P < 0.05 Control vs. AB, †P < 0.05 AB vs. day 1 post AB, ‡P < 0.05 AB vs. day 3 post AB. Note the differences in the scaling of y-axis of the line graphs (A-D and F-G).

Article Snippet: The sections were blocked in Protein Block (Serum-Free, Agilent Dako, #X090930-2) for 10 min and incubated overnight with primary antibodies rabbit anti-mouse versican (GAG-β) antibody (Merck Sigma, Germany, #AB1033, 1:1000), rat anti-human versican antibody (R&D systems, MN, #MAB3054, 1:100), and rabbit anti-mouse/human versican V0, V1 neoepitope antibody (Invitrogen, #PA1-1748A, 1:1000) at 4◦C.

Techniques: Expressing, Control, Western Blot, Staining

Journal: Matrix biology plus

Article Title: Temporal expression and spatial distribution of the proteoglycan versican during cardiac fibrosis development.

doi: 10.1016/j.mbplus.2023.100135

Figure Lengend Snippet: Fig. 4. Cellular origin of versican during development of cardiac fibrosis determined by single molecular fluorescence in situ hybridization (smFISH). Representative confocal images of Col1a1 (red), and Vcan (green) in the heart after sham operation (A-C, Control, n = 3) and aortic banding at day 3 (D-F, AB, n = 3) and at day 56 (G-I). DAPI stain for the nuclei (blue). (F) Versican expression by non-cardiomyocytes and cardiomyocytes after 3 days of aortic banding is shown in pink and orange boxes, respectively. The white arrows show positive signal for Col1a1 (A, D, G), green arrows show positive signal for Vcan (E, H), and yellow arrows show colocalization of Col1a1 and Vcan (F, I). Scale bar = 20 μm, 3 μm (insets). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The sections were blocked in Protein Block (Serum-Free, Agilent Dako, #X090930-2) for 10 min and incubated overnight with primary antibodies rabbit anti-mouse versican (GAG-β) antibody (Merck Sigma, Germany, #AB1033, 1:1000), rat anti-human versican antibody (R&D systems, MN, #MAB3054, 1:100), and rabbit anti-mouse/human versican V0, V1 neoepitope antibody (Invitrogen, #PA1-1748A, 1:1000) at 4◦C.

Techniques: Fluorescence, In Situ Hybridization, Control, Staining, Expressing

Journal: Matrix biology plus

Article Title: Temporal expression and spatial distribution of the proteoglycan versican during cardiac fibrosis development.

doi: 10.1016/j.mbplus.2023.100135

Figure Lengend Snippet: Fig. 6. Versican (VCAN) accumulates in perivascular regions, and in areas of replacement and interstitial fibrosis in aortic banded hearts. (A, B) Representative confocal images of versican (red), and quantification (normalized to perimeter of the vessel, and relative to respective Control) in the cardiac perivascular region of mice after sham (Control, white bar, n = 6) and aortic banding operations (AB, grey bar, n = 6) at day 3, 14 and 56. The white arrows show staining for versican in the tunica adventitia of the vessels, yellow arrows show versican in the surrounding interstitium. (C, D) Representative Masson’s trichrome stained images showing perivascular fibrosis, and quantification (fibrotic area normalized to perimeter of vessel, and relative to respective Control) of Control (white bar, n = 6) and AB hearts (grey bar, n = 6) at day 3, 14 and 56. The yellow arrows show progression of perivascular fibrosis into the surrounding interstitium after aortic banding. Scale bar = 35 μm. (E-G) Representative confocal images of versican and Masson’s trichrome stained images from Control and AB hearts after 56 days showing (F) replacement and (G) interstitial fibrosis. Wheat germ agglutinin (WGA, green) stain for the cardiomyocyte sarcolemma and extracellular matrix, versican (red), and DAPI stain for the nuclei (blue). The white arrows show positive staining for versican and yellow color shows colocalization at fibrotic regions. Scale bar = 15 μm for confocal images, 20 μm for Masson’s trichrome stained images. Data represent mean ± SD. Repeated measures two-way ANOVA with Bonferroni’s multiple com parisons test was used for statistical analysis. P values < 0.05 were considered statistically significant. *P < 0.05 Control vs. AB, ‡P < 0.05 AB vs. day 3 post AB. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The sections were blocked in Protein Block (Serum-Free, Agilent Dako, #X090930-2) for 10 min and incubated overnight with primary antibodies rabbit anti-mouse versican (GAG-β) antibody (Merck Sigma, Germany, #AB1033, 1:1000), rat anti-human versican antibody (R&D systems, MN, #MAB3054, 1:100), and rabbit anti-mouse/human versican V0, V1 neoepitope antibody (Invitrogen, #PA1-1748A, 1:1000) at 4◦C.

Techniques: Control, Staining

Journal: Matrix biology plus

Article Title: Temporal expression and spatial distribution of the proteoglycan versican during cardiac fibrosis development.

doi: 10.1016/j.mbplus.2023.100135

Figure Lengend Snippet: Fig. 7. Production and localization of versican (VCAN) in patients with hypertrophic obstructive cardiomyopathy (HOCM). (A) Relative mRNA expression of VCAN, (B) collagen I alpha 2 (COL1A2), (C) and collagen III alpha 1 (COL3A1) in the left ventricular tissue from patients with HOCM (n = 13) compared to control hearts (Control, n = 4). (D) Correlation (R2) between relative mRNA expression of VCAN and COL1A2, and (E) COL3A1 in HOCM. (F) mRNA expression of versican isoforms V0, V1, V2, V3 in HOCM (n = 13) and Control (n = 4). The values are relative to average of total versican expression in control hearts. (G) Representative confocal images of versican (red) from Control left ventricle tissue and HOCM with (H) interstitial, (I) replacement and (J) perivascular fibrosis. (K) Quantification of versican protein level in HOCM (n = 10) versus Control (n = 4) from the confocal images. Positively stained area for versican was normalized to wheat germ agglutinin (WGA, green) stain for cardiomyocyte sarcolemma and extracellular matrix. Values relative to the average of Control are reported. Data points in the bar graph (K) represent the average value from n = 8 images per sample. DAPI stain for the nuclei (blue). White arrows show positive staining for versican and yellow color shows colocalization at fibrotic regions. Scale bar = 15 μm. Bar graph with scatter plot shows mean ± SD. Welch’s t-test (A, B, C, F), Student’s t-test (K), and Pearson correlation coefficient (D, E) were used for statistical analysis. P values < 0.05 were considered statistically significant. *P < 0.05, **P < 0.01, ***P < 0.001 Control vs. HOCM. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The sections were blocked in Protein Block (Serum-Free, Agilent Dako, #X090930-2) for 10 min and incubated overnight with primary antibodies rabbit anti-mouse versican (GAG-β) antibody (Merck Sigma, Germany, #AB1033, 1:1000), rat anti-human versican antibody (R&D systems, MN, #MAB3054, 1:100), and rabbit anti-mouse/human versican V0, V1 neoepitope antibody (Invitrogen, #PA1-1748A, 1:1000) at 4◦C.

Techniques: Expressing, Control, Staining

Journal: Matrix biology plus

Article Title: Temporal expression and spatial distribution of the proteoglycan versican during cardiac fibrosis development.

doi: 10.1016/j.mbplus.2023.100135

Figure Lengend Snippet: Fig. 8. Expression and localization of versican (VCAN) in patients with dilated cardiomyopathy (DCM). (A) Relative mRNA expression of VCAN, (B) collagen I alpha 2 (COL1A2), (C) collagen III alpha 1 (COL3A1) in left ventricular tissue from patients with DCM (n = 11) compared to control hearts (Control, n = 4). (D) Correlation (R2) between relative mRNA expression of VCAN and COL1A2, and (E) COL3A1 in DCM. (F) mRNA expression of versican isoforms V0, V1, V2, V3 in DCM (n = 11) and Control (n = 4). The values are relative to average of total versican expression in control hearts. (G) Representative confocal images of versican (red) from Control left ventricle tissue, and DCM with (H) interstitial, (I) replacement and (J) perivascular fibrosis. (K) Quantification of versican protein level in DCM (n = 10) versus Control (n = 4) from the confocal images. Positively stained area for versican was normalized to wheat germ agglutinin (WGA, green) stain for cardiomyocyte sarcolemma and extracellular matrix. Values relative to the average of Control are reported. Data points in the bar graph (K) represent the average value from n = 8 images per sample. DAPI stain for the nuclei (blue). White arrows show positive staining for versican and yellow color shows colocalization at fibrotic regions. Scale bar = 15 μm. Bar graph with scatter plot shows mean ± SD. Welch’s t-test (A, B, C, F), Student’s t-test (K), and Pearson correlation coefficient (D, E) were used for statistical analysis. P values < 0.05 were considered statistically significant. *P < 0.05, **P < 0.01 Control vs. DCM. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The sections were blocked in Protein Block (Serum-Free, Agilent Dako, #X090930-2) for 10 min and incubated overnight with primary antibodies rabbit anti-mouse versican (GAG-β) antibody (Merck Sigma, Germany, #AB1033, 1:1000), rat anti-human versican antibody (R&D systems, MN, #MAB3054, 1:100), and rabbit anti-mouse/human versican V0, V1 neoepitope antibody (Invitrogen, #PA1-1748A, 1:1000) at 4◦C.

Techniques: Expressing, Control, Staining

Journal: Cancers

Article Title: Non-Oncogene Addiction of KRAS -Mutant Cancers to IL-1β via Versican and Mononuclear IKKβ.

doi: 10.3390/cancers15061866

Figure Lengend Snippet: Figure 4. Tumor-secreted versican drives macrophage IKKβ, metastasis, and is a cancer biomarker. (A–E) Kras-mutant and wild-type cancer cell RNA and supernatants were subjected to microar- ray and LC-MSMS analyses, respectively. Shown are the experimental design (A), top-20 over- represented transcripts (B) and secretory proteins (C), and Vcan/VCAN mRNA/protein expression (D,E). n = 2–3/group; P, probability, two-way ANOVA; Bold letters, false discovery rate (FDR) q < 0.05 compared with Kras-wild-type cells, two-stage linear step-up procedure of Benjamini, Hochberg, and Yekutieli. (F,G) Representative bioluminescent images (F) and data summary (G) from pNGL RAW264.7 cells exposed to lipopolysaccharide (LPS; 1 µg/mL) or recombinant proteins (1–2 nM). n = 5/group; P, probability, two-way ANOVA; ****, p < 0.0001 compared with other groups, Bonferroni post-tests. (H) Immunoblots of mouse BMDM exposed to VCAN. n = 5/group. (I–M) LLC cells stably expressing

Article Snippet: Control shRNA (shC, sc-108080-V; target sequences are proprietary of the manufacturer) and anti-mouse Vcan shRNA (shVcan, sc-41904–V) pools were from Santa Cruz.

Techniques: Biomarker Discovery, Mutagenesis, Expressing, Recombinant, Western Blot, Stable Transfection

Journal: PLoS ONE

Article Title: Versican is differentially regulated in the adventitial and medial layers of human vein grafts

doi: 10.1371/journal.pone.0204045

Figure Lengend Snippet: Antibodies and reagents used in this study.

Article Snippet: Proteins were transferred to nitrocellulose and probed with anti-human versican (0.1 μg/ml AF3054 [R&D systems, Minneapolis, MN] or 0.06 μg/ml 2B1 [270428 Seikagaku] for cultured cells and tissue, respectively).

Techniques: Concentration Assay, Binding Assay, Negative Control

Journal: PLoS ONE

Article Title: Versican is differentially regulated in the adventitial and medial layers of human vein grafts

doi: 10.1371/journal.pone.0204045

Figure Lengend Snippet: (A) Versican immunostaining in pre-culture and 14 day cultured floating vein rings was quantified for the intima/media and adventitia (B). The dotted line indicates the adventitial/medial boundary (* P = 0.01 day 0 vs day 14; N = 19 veins). (C) Versikine immunostaining in pre-culture and 14 day cultured floating vein rings was quantified (D) for the intima/media and adventitia (*P<0.04; **P<0.0001 day 0 vs day 14; N = 19 veins). Scale bars indicate 500 μm.

Article Snippet: Proteins were transferred to nitrocellulose and probed with anti-human versican (0.1 μg/ml AF3054 [R&D systems, Minneapolis, MN] or 0.06 μg/ml 2B1 [270428 Seikagaku] for cultured cells and tissue, respectively).

Techniques: Immunostaining, Cell Culture

Journal: PLoS ONE

Article Title: Versican is differentially regulated in the adventitial and medial layers of human vein grafts

doi: 10.1371/journal.pone.0204045

Figure Lengend Snippet: Induction of versican message and protein in floating intimal/medial and adventitial tissue. (A) Versican mRNA (*P<0.02 intima/media vs. adventitia. N = 3 veins day 0, N = 6 veins days 1–7). (B) Versican western blot of tissue from one vein (“int/med” intima/media and “adv” adventitia. (C) Quantification of V0/V1 versican protein (*P<0.02 intima/media vs. adventitia mean gray value. N = 3 veins). (D) Quantification of versican degradation products (75–250 kD) relative to intact V0/V1 versican (N = 3 veins).

Article Snippet: Proteins were transferred to nitrocellulose and probed with anti-human versican (0.1 μg/ml AF3054 [R&D systems, Minneapolis, MN] or 0.06 μg/ml 2B1 [270428 Seikagaku] for cultured cells and tissue, respectively).

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Versican is differentially regulated in the adventitial and medial layers of human vein grafts

doi: 10.1371/journal.pone.0204045

Figure Lengend Snippet: Expression of inhibitors of versican-degrading proteases in cultured adventitial and intimal/medial tissue.

Article Snippet: Proteins were transferred to nitrocellulose and probed with anti-human versican (0.1 μg/ml AF3054 [R&D systems, Minneapolis, MN] or 0.06 μg/ml 2B1 [270428 Seikagaku] for cultured cells and tissue, respectively).

Techniques: Expressing, Cell Culture

Journal: PLoS ONE

Article Title: Versican is differentially regulated in the adventitial and medial layers of human vein grafts

doi: 10.1371/journal.pone.0204045

Figure Lengend Snippet: Histochemical and immunostaining of neointimal cells and capsule cells (left and right panel, respectively) for (A) versican, (B) hyaluronan, (C) versikine, and (D) ADAMTS4. * indicates the adventitial surface. Scale bars indicate 100 μm.

Article Snippet: Proteins were transferred to nitrocellulose and probed with anti-human versican (0.1 μg/ml AF3054 [R&D systems, Minneapolis, MN] or 0.06 μg/ml 2B1 [270428 Seikagaku] for cultured cells and tissue, respectively).

Techniques: Immunostaining

Journal: PLoS ONE

Article Title: Versican is differentially regulated in the adventitial and medial layers of human vein grafts

doi: 10.1371/journal.pone.0204045

Figure Lengend Snippet: (A) The left panel shows an adventitial vessel in cross and tangential section stained for CD31 (black: endothelium) and CD34 (red: CD34 + progenitor cell and endothelium but the latter is masked by the black stain) and the right panel shows a cross and tangential section of a vessel stained for CD34 (red: CD34 + progenitor cell and endothelium) and SMA (black: pericytes). (B) Vessels stained for versican and hyaluronan (brown) in the top and bottom panels, respectively; and (C) a vessel stained for CD34 (red) and versican (black). The arrowhead indicates CD34 and versican co-localization compared to the arrow that indicates unlabeled pericytes. Scale bars indicate 100 μm in A and B, and 50 μm in C.

Article Snippet: Proteins were transferred to nitrocellulose and probed with anti-human versican (0.1 μg/ml AF3054 [R&D systems, Minneapolis, MN] or 0.06 μg/ml 2B1 [270428 Seikagaku] for cultured cells and tissue, respectively).

Techniques: Staining

Journal: PLoS ONE

Article Title: Versican is differentially regulated in the adventitial and medial layers of human vein grafts

doi: 10.1371/journal.pone.0204045

Figure Lengend Snippet: Versican changes in different ex vivo vein graft models.

Article Snippet: Proteins were transferred to nitrocellulose and probed with anti-human versican (0.1 μg/ml AF3054 [R&D systems, Minneapolis, MN] or 0.06 μg/ml 2B1 [270428 Seikagaku] for cultured cells and tissue, respectively).

Techniques: Ex Vivo

Journal: PLoS ONE

Article Title: Versican is differentially regulated in the adventitial and medial layers of human vein grafts

doi: 10.1371/journal.pone.0204045

Figure Lengend Snippet: Cells were stimulated with either 10% FBS (A) or 10 ng/ml PDGF-BB (B). Adventitial cells had significantly higher induction of versican, HAS1 and HAS2 compared to SMCs (***P<0.0001; **P<0.01; *P<0.05). Data are from pairs of cells from 3 (A) and 10 (B) veins in duplicate.

Article Snippet: Proteins were transferred to nitrocellulose and probed with anti-human versican (0.1 μg/ml AF3054 [R&D systems, Minneapolis, MN] or 0.06 μg/ml 2B1 [270428 Seikagaku] for cultured cells and tissue, respectively).

Techniques:

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Versican: a novel modulator of hepatic fibrosis.

doi: 10.1038/labinvest.2015.152

Figure Lengend Snippet: Figure 1 Versican expression in human liver cirrhosis and mouse liver fibrosis. (a) Sections of liver parenchyma from non-cirrhotic (n = 5) and cirrhotic (n = 5) human livers were stained using immunohistochemistry for versican (12C5), alongside normal (n = 5) and fibrotic (n = 5) mouse liver sections (after 4 weeks CCl4) (HPA004726). Representative non-cirrhotic human liver shown from a patient with metastatic colorectal carcinoma and representative cirrhotic human liver shown from a patient with primary sclerosing cholangitis. (b) Quantitation of versican immunostaining was performed with image thresholding and significance determined by two-way analysis of variance (*) (Po0.05). Mean value per lane is shown (n = 5) and error bars represent the s.e.m. CCL4, carbon tetrachloride.

Article Snippet: Data normalization relative to the 18 s RNA subunit was calculated using the comparative Ct (ΔΔCt) method to calculate the fold change between the control and experimental groups as previously described.77 Primer pairs used for the different mouse and human genes are listed in Table 1. siRNA Constructs and Cell Transfections The following expression plasmids were used: human versican siRNA (Santa Cruz cat. # sc-41903) and control siRNA (Santa Cruz cat. # sc-37007).

Techniques: Expressing, Staining, Immunohistochemistry, Quantitation Assay, Immunostaining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Versican: a novel modulator of hepatic fibrosis.

doi: 10.1038/labinvest.2015.152

Figure Lengend Snippet: Figure 3 Versican expression increases during liver fibrosis and reduces during recovery. (a) Total RNA was extracted from the livers of control mice, CCl4 4-week treated mice and mice recovering from CCl4-induced liver injury and analyzed by qPCR for versican and collagen-1 mRNA expression using 18 s RNA as endogenous control. (b) Liver injury and fibrogenesis were assessed in control mice for TNF-α, α-SMA, and TGF-β. Relative mRNA expression and s.e.m. values are shown. Significant differences determined with one-way analysis of variance are indicated compared with control (*) and compared with 4 weeks of treatment (†) (Po0.05, n = 9–10). CCL4, carbon tetrachloride; α-SMA, α-smooth muscle actin; TNF-α, tumor necrosis factor-α; TGF-β, transforming growth factor-β.

Article Snippet: Data normalization relative to the 18 s RNA subunit was calculated using the comparative Ct (ΔΔCt) method to calculate the fold change between the control and experimental groups as previously described.77 Primer pairs used for the different mouse and human genes are listed in Table 1. siRNA Constructs and Cell Transfections The following expression plasmids were used: human versican siRNA (Santa Cruz cat. # sc-41903) and control siRNA (Santa Cruz cat. # sc-37007).

Techniques: Expressing, Control

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Versican: a novel modulator of hepatic fibrosis.

doi: 10.1038/labinvest.2015.152

Figure Lengend Snippet: Figure 4 CCl4-induced liver fibrosis leads to increase in α-SMA and versican V0 fragment. Total protein was extracted from liver samples of control, CCl4 4-week treated and CCl4-treated recovering mice. (a) Total protein samples were analyzed for mouse α-SMA expression using histone H3 as a protein normalizing control. (b and c) Total protein samples were analyzed for the ADAMTS generated versican fragments by western blot analysis using anti- DPEAAE neo-epitope antibody and β-actin as a protein normalizing control. Significant differences determined with one-way analysis of variance are indicated compared with control (*) and compared with 4 weeks of treatment (†) (Po0.05). Western blots shown demonstrate three representative lanes per time point. Data are representative of 3–4 repeat experiments. Mean band densitometry values per lane are shown (n = 9–10) and error bars represent the s.e.m. ADAMTS, A Disintegrin-like and Metalloproteinase with Thrombospondin-1 motifs; CCL4, carbon tetrachloride; α-SMA, α-smooth muscle actin.

Article Snippet: Data normalization relative to the 18 s RNA subunit was calculated using the comparative Ct (ΔΔCt) method to calculate the fold change between the control and experimental groups as previously described.77 Primer pairs used for the different mouse and human genes are listed in Table 1. siRNA Constructs and Cell Transfections The following expression plasmids were used: human versican siRNA (Santa Cruz cat. # sc-41903) and control siRNA (Santa Cruz cat. # sc-37007).

Techniques: Control, Expressing, Generated, Western Blot

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Versican: a novel modulator of hepatic fibrosis.

doi: 10.1038/labinvest.2015.152

Figure Lengend Snippet: Figure 6 Activation of primary mouse stellate cells leads to increased versican expression. Primary HSCs were isolated from 3- to 6-week-old C57BL/6 mice as indicated in the methods. Isolated primary HSCs were activated by plastic adherence over 1, 3, 5, 7, 14, and 21 days. Total RNA was then extracted from cells and analyzed for (a) versican, (b) collagen-1, and (c) α-SMA mRNA expression using 18 s RNA as an endogenous control. (d) ADAMTS-1 and -4 were analyzed alongside TIMP-1 and -3 expression levels, using 18 S RNA as an endogenous control. Significant differences determined with one-way analysis of variance are indicated compared with day 1 (*) (Po0.05). Data are representative of 3–4 repeat experiments. Mean value per lane is shown (n = 3) and error bars represent the s.e.m. ADAMTS, A Disintegrin-like and Metalloproteinase with Thrombospondin-1 motifs; HSC, hepatic stellate cell; α-SMA, α-smooth muscle actin; TIMP-1, tissue inhibitor of metalloproteinase-1.

Article Snippet: Data normalization relative to the 18 s RNA subunit was calculated using the comparative Ct (ΔΔCt) method to calculate the fold change between the control and experimental groups as previously described.77 Primer pairs used for the different mouse and human genes are listed in Table 1. siRNA Constructs and Cell Transfections The following expression plasmids were used: human versican siRNA (Santa Cruz cat. # sc-41903) and control siRNA (Santa Cruz cat. # sc-37007).

Techniques: Activation Assay, Expressing, Isolation, Control

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Versican: a novel modulator of hepatic fibrosis.

doi: 10.1038/labinvest.2015.152

Figure Lengend Snippet: Figure 7 Transient knockdown of versican in LX2 cells leads to a decrease in versican V1 fragment and decreased HSC activation. Total RNA and protein were extracted from LX2 cells 48 h after transient knockdown of versican or not as indicated. Total RNA was analyzed for (a) collagen-1, (b) α-SMA, (c) KI67, and (d) TGF-β1 mRNA expression, respectively, by qPCR using 18 s RNA as an endogenous control. (e) Total protein extracts were analyzed for versican V1 fragment using β-actin as a protein normalizing control. (f) Total protein was extracted from LX2 cells after versican KD, staurosporine treatment or not as indicated and subjected to western blot analysis for PARP as indicated. (g) Total RNA was extracted from primary mouse HSC cultures 48 h after knockdown of versican or not (control) as indicated. mRNA expression was analyzed by qPCR for versican, α-SMA, collagen-1, and TGF-β1 using 18 s RNA as an endogenous control. Significant differences determined with one-way analysis of variance are indicated compared with no treatment control (*) (Po0.05). Data are representative of 3–4 repeat experiments. The mean band densitometry value per lane is shown (n = 3) in (e) and error bars represent the s.e.m. HSC, hepatic stellate cell; α-SMA, α-smooth muscle actin; TGF-β, transforming growth factor-β; PARP, poly(ADP-ribose)polymerase.

Article Snippet: Data normalization relative to the 18 s RNA subunit was calculated using the comparative Ct (ΔΔCt) method to calculate the fold change between the control and experimental groups as previously described.77 Primer pairs used for the different mouse and human genes are listed in Table 1. siRNA Constructs and Cell Transfections The following expression plasmids were used: human versican siRNA (Santa Cruz cat. # sc-41903) and control siRNA (Santa Cruz cat. # sc-37007).

Techniques: Knockdown, Activation Assay, Expressing, Control, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Screening and identification of versican as a sensitive biomarker and potential therapeutic target in basal cell carcinoma

doi: 10.7150/ijms.105650

Figure Lengend Snippet: Silencing of VCAN regulates A431 cells proliferation, migration, invasion, and apoptosis. A: Cell migration rate was observed by wound healing assay at 24h, 48h and 72h; B: CCK-8 was used to detect the effect of VCAN on A431 cells proliferation under different treatments. C: 24h migration rate of A431(Top: Nci; Bottom: si-VCAN); D-I: Samples were collected from A431 cells after DOX treatment for 24h post-transfection, and the expression levels of total BAX, BCL-2, c-Caspase3, PTEN, phosphorylated and total mTOR, DDX5 and GAPDH were examined by Western blotting (n=3). J, K : Representative images of TUNEL and Hoechst 33342 staining of A431 cells (n=5). ns: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Control lentiviral particles (sc-437282) and VCAN shRNA lentiviral particles (sc-41903-V) were purchased from Santa Cruz Biotechnology.

Techniques: Migration, Wound Healing Assay, CCK-8 Assay, Transfection, Expressing, Western Blot, TUNEL Assay, Staining

Journal: International Journal of Medical Sciences

Article Title: Screening and identification of versican as a sensitive biomarker and potential therapeutic target in basal cell carcinoma

doi: 10.7150/ijms.105650

Figure Lengend Snippet: A-C: Nude mice were injected subcutaneously in the axillary region with 5×10 6 control cells (sh-NC) or a VCAN knockout cell (sh-VCAN) for two weeks. Tumours were dissected and tumour/body weight calculated (n=6). Scale bar: 100 µm. D, E : Immunofluorescence of the macrophage M1 polarization marker iNOS (n=3). Scale bar: 100 µm. F : Immunohistochemical staining was used to analyse the expression levels of VCAN in normal human skin and tumour tissue from BCC patients (n=3). Scale bar: 50 µm. ns: not significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Control lentiviral particles (sc-437282) and VCAN shRNA lentiviral particles (sc-41903-V) were purchased from Santa Cruz Biotechnology.

Techniques: Injection, Control, Knock-Out, Immunofluorescence, Marker, Immunohistochemical staining, Staining, Expressing