vimentin Search Results


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  • 97
    Millipore vimentin vim
    Evidence of epithelial to mesenchymal transition (EMT) in chronic arsenic treated CATLE cells. (A) Quantitative expression of EMT markers including loss of CDH1 protein and increased <t>VIM</t> protein in CATLE cells (lower figure). Quantitative protein data
    Vimentin Vim, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    PhosphoSolutions vimentin
    Evidence of epithelial to mesenchymal transition (EMT) in chronic arsenic treated CATLE cells. (A) Quantitative expression of EMT markers including loss of CDH1 protein and increased <t>VIM</t> protein in CATLE cells (lower figure). Quantitative protein data
    Vimentin, supplied by PhosphoSolutions, used in various techniques. Bioz Stars score: 88/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Agilent technologies vimentin vim
    CD133 + cells in IC-1425EPN xenograft tumors. (A) Representative graphs showing the isolation of CD133 + cells from an IC-1425EPN xenograft tumor (passage II) with FACS using PE-conjugated monoclonal antibodies against human CD133. (B) In vitro formation of neurosphere from isolated CD133 + cells in the presence of EGF and bFGF (upper panel), and induced differentiation of preformed neurosphere with FBS (lower panel). (C and D) Representative IMF images showing the expression of human-specific mitochondrial (MT) antigen, markers of neuroprogenitor (Nestin), early (TuJ-1), and mature (MAP2 and SYP) neurons, glial precursor (A2B5), astrocyte (GFAP), as well as intermediate filament <t>VIM</t> (×60).
    Vimentin Vim, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 90/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Cell Signaling Technology Inc vimentin vim
    MTUS1/ATIP3a overexpression is related to the migration and invasion of SACC. (A) Differential expression of ERK-Slug pathway in SACC-LM cells treated with either control vector or ATIP3a plasmid. SACC-LM cells displayed decreased Slug, <t>Vimentin</t> and pERK1/2 protein levels and increased E-cadherin, ATIP3a protein levels upon ATIP3a overexpression. ATIP3a protein was detected by anti-MTUS1 antibody. ( B and C ) The migration and invasion ability of SACC cells was assessed by a transwell migration and invasion assay. ATIP3a overexpression significantly inhibited the migration (B) and invasion (C) of SACC-LM cells. (D) Cell proliferation was measured using a MTT assay. The cell proliferation rate of SACC-LM was significantly inhibited after overexpressed with MTUS1/ATIP3a. *: P
    Vimentin Vim, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    ABclonal vimentin vim
    The influences of CAV3.1 knockdown on expressions of proteins involving cell cycle, EMT, and Akt activity in PCa cells. Notes: ( A ) CAV3.1 knockdown led to decreased CCND1, but not for CCNE1 and CCNB1 in PC-3 and DU145 cells. Moreover, CAV3.1 knockdown led to decreased N-cad and <t>Vim,</t> 2 mesenchymal markers, and increased E-cad, an epithelial marker. ( B ) CAV3.1 knockdown led to decreased p-Akt (S473) and p-Akt (T308) in PC-3 and DU145 cells. Abbreviations: E-cad, E-cadherin; EMT, epithelial–mesenchymal transition; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; N-cad, N-cadherin; PCa, prostate cancer.
    Vimentin Vim, supplied by ABclonal, used in various techniques. Bioz Stars score: 93/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Boehringer Mannheim anti vimentin vim
    The influences of CAV3.1 knockdown on expressions of proteins involving cell cycle, EMT, and Akt activity in PCa cells. Notes: ( A ) CAV3.1 knockdown led to decreased CCND1, but not for CCNE1 and CCNB1 in PC-3 and DU145 cells. Moreover, CAV3.1 knockdown led to decreased N-cad and <t>Vim,</t> 2 mesenchymal markers, and increased E-cad, an epithelial marker. ( B ) CAV3.1 knockdown led to decreased p-Akt (S473) and p-Akt (T308) in PC-3 and DU145 cells. Abbreviations: E-cad, E-cadherin; EMT, epithelial–mesenchymal transition; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; N-cad, N-cadherin; PCa, prostate cancer.
    Anti Vimentin Vim, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abnova anti vimentin vim
    HuCCT1 and Huh28 cell lines exhibit an epithelioid and a mesenchymal‐like phenotype respectively, the later correlating with endogenous activation of the TGFβ pathway. (A) Phase contrast micrographs of HuCCT1 and Huh28 cells (left panel; scale bar, 100 µm) and expression of <t>vimentin</t> and E‐cadherin at the RNA (middle panel) and protein level (right panel). (B) Volcano plot of genes differentially expressed between HuCCT1 and Huh28 (left panel) and GSEA highlighting a TGFβ signature enriched in the gene expression profile of Huh28. (C) Enzyme‐linked immunosorbent assay demonstrating increased secretion of TGFβ1 in the supernatant of Huh28 cells 24 hours after overnight serum starvation. (D) Relative luciferase activity of HuCCT1 and Huh28 engineered cell lines in which a luciferase gene is under the transcriptional control of a TATA box alone (white bars) or with SMAD responsive elements (black bars). Luciferase activity was measured at the basal level without exogenous addition of TGFβ after overnight serum starvation. (E) Gene expression analysis of TGFβ target genes in HuCCT1 and Huh28 cells at the basal level by microarray and Q‐RT‐PCR. (F) Q‐RT‐PCR analysis of TGFβ target gene SMAD7 in Huh28 cells treated with 10 µM TGFβ inhibitors (black bars). Statistical analysis was performed by t test (* P
    Anti Vimentin Vim, supplied by Abnova, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson vim vimentin
    BHLHE41 expression suppressed tumour cell migration and invasion of MCF‐7 cells and MDA‐MB‐231 cells. A, B, The mRNA levels of CLDN1, CLDN4 and CDH1 were up‐regulated, whereas those of SNAI1, SNAI2, <t>VIM</t> and CDH2 were down‐regulated after transfecting with exogenous BHLHE41 in MCF‐7 and MDA‐MB‐231 cells. C, D, WB analysis showed comparable results in the expression of the above genes. E‐H, Transwell assay for assessing cell migration and invasion of MCF‐7 and MDA‐MB‐231. As measured by the transwell assay without or with gel cover, BHLHE41 overexpression led to a significant decrease in the number of migrated and invasion cells. Transwell assays were repeated in triplicate. Ten views were selected for each trial, and the migrated or invasion cells were counted for the final statistical analysis. Each value represents the mean ± SE (bars) of three independent experiments, *** P
    Vim Vimentin, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology vimentin vim
    In vitro characterization of pituitary adenoma cell cultures for mesenchymal stromal cell markers. Representative FACS histograms of pituitary somatotroph adenoma cell culture (a–f). Cells were positive for CD90 (a), CD105 (b), and CD44 (c) and were negative for CD34 (d), CD45 (e), and HLA-DR (f). Representative immunostaining of <t>VIM</t> (green) (g) and negative control (h). Nuclei were stained with DAPI (blue). 200x magnification. CD90, Thy-1 cell surface antigen; CD105, endoglin; CD44, CD44 molecule (Indian blood group); CD34, CD34 molecule; CD45, protein tyrosine phosphatase, receptor type, C; HLA-DR, major histocompatibility complex, class II, DR.
    Vimentin Vim, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc anti vimentin vim
    Western blot showing APD-induced changes in the expression of the HIST1H4B, HSPA8, NCL, NPM1, and <t>VIM</t> proteins in C6 and B35 cells and the rat cortex. Nuclear protein extracts collected from APD-treated C6 cells, B35 cells and rat cortex were compared. a and ( b ), HIST1H4B; ( c ) and ( d ),HSPA8; ( e ) and ( f ), NCL; ( g ) and ( h ), NPM1; ( i ) and ( j ), VIM. Bar charts were constructed from triplicate western blot data obtained from three different batches of APD-treated cells or rat cortices using ANOVA followed by Dunnett’s test (* p -value
    Anti Vimentin Vim, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam vimentin
    TGF-β1 induced EMT progress and restored migration, invasion abilities suppressed by Neferine. (a,b) TGF-β1 induced EMT progress. HCC cells were treated with 5 ng/ml, 10 ng/ml, 20 ng/ml TGF-β1 for 24 hrs or 48 hrs to induce EMT. mRNA and protein expression of EMT biomarkers (E-cadherin, N-cadherin <t>Vimentin),</t> and EMT promoting transcription factor (Snail) was determined by qRT-PCR and by Western blot. Original blots of high-contrast blots are presented in Supplementary Fig. S4 . (c) Morphological changes of HCC cells treated with 10 ng/ml TGF-β1 for different times. TGF-β1 changed cells morphology from pebble-like epithelial to dispersed, spindle-like mesenchymal and pseudopodium stretching. (Original magnification: ×400).
    Vimentin, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 7685 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Abcam vimentin antibody
    GATA3 upregulates E-cadherin and downregulates the expression of N-cadherin and <t>vimentin</t> in OS cells. Notes: ( A , B ) GATA3 was overexpressed in U2OS and Saos2 cells. The expression of EMT markers, such as epithelial marker (E-cadherin) and mesenchymal markers (N-cadherin and vimentin), were detected by qRT-PCR and western blotting assay. *
    Vimentin Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 98/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore α vimentin
    Soft substrate induces a rapid and homogeneous epithelial differentiation of RLSCs. (a) Phase-contrast micrographs of RLSCs grown on Petri plastic dish (CTRL;  E >  1 GPa) and on hydrogels with  E  = 0.4 kPa and 80 kPa, for 24 and 48 hours. Images are representative of three independent experiments. Scale bar: 100  μ m. (b) Phase-contrast micrographs and immunofluorescence of cells cultured on plastic (CTRL), 0.4 kPa and 80 kPa for 24 and 48 hours, stained for HNF4 α , Vimentin, and E-cadherin. The nuclei were stained with DAPI. Images are representative of three independent experiments. Scale bar: 50  μ m. (c) Western blot analysis of Vimentin at 24 and 48 hours after seeding on substrates with the indicated  E  values. CDK4 was used as a loading control.
    α Vimentin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Syntaxin vimentin vim
    Soft substrate induces a rapid and homogeneous epithelial differentiation of RLSCs. (a) Phase-contrast micrographs of RLSCs grown on Petri plastic dish (CTRL;  E >  1 GPa) and on hydrogels with  E  = 0.4 kPa and 80 kPa, for 24 and 48 hours. Images are representative of three independent experiments. Scale bar: 100  μ m. (b) Phase-contrast micrographs and immunofluorescence of cells cultured on plastic (CTRL), 0.4 kPa and 80 kPa for 24 and 48 hours, stained for HNF4 α , Vimentin, and E-cadherin. The nuclei were stained with DAPI. Images are representative of three independent experiments. Scale bar: 50  μ m. (c) Western blot analysis of Vimentin at 24 and 48 hours after seeding on substrates with the indicated  E  values. CDK4 was used as a loading control.
    Vimentin Vim, supplied by Syntaxin, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Evidence of epithelial to mesenchymal transition (EMT) in chronic arsenic treated CATLE cells. (A) Quantitative expression of EMT markers including loss of CDH1 protein and increased VIM protein in CATLE cells (lower figure). Quantitative protein data

    Journal: Toxicology and applied pharmacology

    Article Title: Chronic Inorganic Arsenic Exposure In Vitro Induces a Cancer Cell Phenotype in Human Peripheral Lung Epithelial Cells

    doi: 10.1016/j.taap.2015.03.014

    Figure Lengend Snippet: Evidence of epithelial to mesenchymal transition (EMT) in chronic arsenic treated CATLE cells. (A) Quantitative expression of EMT markers including loss of CDH1 protein and increased VIM protein in CATLE cells (lower figure). Quantitative protein data

    Article Snippet: Antibodies for vimentin (VIM) and β-Actin were obtained from Sigma Chemical Co. (St. Louis, MO).

    Techniques: Expressing

    Representative immunoistochemical images of E-cadherin and vimentin in lungs. Representative immunoistochemical images of E-cadherin and vimentin in lung sections from mice exposed at 30 days to MWCNTs through pharyngeal aspiration. E-cadherin was mainly expressed in type II pneumocytes in both control and MWCNT exposed lungs. The number of E-cadherin positive cells in NP-exposed compared to control sections was decreased ( a , c , e ), while the expression for vimentin was increased ( b , d , f ) (20×; scale bar = 60 μm)

    Journal: Particle and Fibre Toxicology

    Article Title: Multi-walled carbon nanotubes directly induce epithelial-mesenchymal transition in human bronchial epithelial cells via the TGF-β-mediated Akt/GSK-3β/SNAIL-1 signalling pathway

    doi: 10.1186/s12989-016-0138-4

    Figure Lengend Snippet: Representative immunoistochemical images of E-cadherin and vimentin in lungs. Representative immunoistochemical images of E-cadherin and vimentin in lung sections from mice exposed at 30 days to MWCNTs through pharyngeal aspiration. E-cadherin was mainly expressed in type II pneumocytes in both control and MWCNT exposed lungs. The number of E-cadherin positive cells in NP-exposed compared to control sections was decreased ( a , c , e ), while the expression for vimentin was increased ( b , d , f ) (20×; scale bar = 60 μm)

    Article Snippet: Antibodies Primary antibodies to E-cadherin, GAPDH, fibronectin, SNAIL-1 and TBP were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA); primary antibody to β-catenin was from Abcam (Cambridge, UK); primary antibody to vimentin was from Sigma; primary antibody to α-SMA was from GeneTex (Irving, CA).

    Techniques: Mouse Assay, Expressing

    Relative expression of epithelial and mesenchymal markers of BEAS-2B cells exposed to MWCNT. Relative expression of epithelial and mesenchymal markers were checked by qRT-PCR ( a ), WB ( b ), confocal microscopy ( c ) in BEAS-2B cells incubated for 96 h in either the absence (0 μg/ml, control) or presence of different concentrations of pMWCNT or MWCNTg. a Relative expression of E-cadherin, α-SMA and vimentin mRNA. Results were expressed in units of relative mRNA expression compared to control cells ( n = 3; white bar = pMWCNT; black bar = MWCNTg). Versus control * p

    Journal: Particle and Fibre Toxicology

    Article Title: Multi-walled carbon nanotubes directly induce epithelial-mesenchymal transition in human bronchial epithelial cells via the TGF-β-mediated Akt/GSK-3β/SNAIL-1 signalling pathway

    doi: 10.1186/s12989-016-0138-4

    Figure Lengend Snippet: Relative expression of epithelial and mesenchymal markers of BEAS-2B cells exposed to MWCNT. Relative expression of epithelial and mesenchymal markers were checked by qRT-PCR ( a ), WB ( b ), confocal microscopy ( c ) in BEAS-2B cells incubated for 96 h in either the absence (0 μg/ml, control) or presence of different concentrations of pMWCNT or MWCNTg. a Relative expression of E-cadherin, α-SMA and vimentin mRNA. Results were expressed in units of relative mRNA expression compared to control cells ( n = 3; white bar = pMWCNT; black bar = MWCNTg). Versus control * p

    Article Snippet: Antibodies Primary antibodies to E-cadherin, GAPDH, fibronectin, SNAIL-1 and TBP were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA); primary antibody to β-catenin was from Abcam (Cambridge, UK); primary antibody to vimentin was from Sigma; primary antibody to α-SMA was from GeneTex (Irving, CA).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Confocal Microscopy, Incubation

    CD133 + cells in IC-1425EPN xenograft tumors. (A) Representative graphs showing the isolation of CD133 + cells from an IC-1425EPN xenograft tumor (passage II) with FACS using PE-conjugated monoclonal antibodies against human CD133. (B) In vitro formation of neurosphere from isolated CD133 + cells in the presence of EGF and bFGF (upper panel), and induced differentiation of preformed neurosphere with FBS (lower panel). (C and D) Representative IMF images showing the expression of human-specific mitochondrial (MT) antigen, markers of neuroprogenitor (Nestin), early (TuJ-1), and mature (MAP2 and SYP) neurons, glial precursor (A2B5), astrocyte (GFAP), as well as intermediate filament VIM (×60).

    Journal: Neuro-Oncology

    Article Title: A clinically relevant orthotopic xenograft model of ependymoma that maintains the genomic signature of the primary tumor and preserves cancer stem cells in vivo

    doi: 10.1093/neuonc/nop056

    Figure Lengend Snippet: CD133 + cells in IC-1425EPN xenograft tumors. (A) Representative graphs showing the isolation of CD133 + cells from an IC-1425EPN xenograft tumor (passage II) with FACS using PE-conjugated monoclonal antibodies against human CD133. (B) In vitro formation of neurosphere from isolated CD133 + cells in the presence of EGF and bFGF (upper panel), and induced differentiation of preformed neurosphere with FBS (lower panel). (C and D) Representative IMF images showing the expression of human-specific mitochondrial (MT) antigen, markers of neuroprogenitor (Nestin), early (TuJ-1), and mature (MAP2 and SYP) neurons, glial precursor (A2B5), astrocyte (GFAP), as well as intermediate filament VIM (×60).

    Article Snippet: Primary antibodies included the following: mouse antisynaptophysin (anti-SYP) (1:200), glial fibrillary acidic protein (GFAP) (1:200), vimentin (VIM) (1:200), epithelial membrane antigen (EMA) (1:100), α-smooth muscle actin (SMA) (1:100) (Dako), MAP-2 (1:200), Ki-67 (1:20) (Abcam, Inc.), neuron-specific class III B-tublin (TuJ-1) (1:100) (Fitzgerald Industries International, Inc.), human-specific mitochondria (MT) (1:50) (Chemicon International, Inc.); VEGF (c-1) (1:100) (Santa Cruz Biotechnology, Inc.); and rabbit anti-Nestin (NES) (1:500), anti-Von Willebrand Factor (vWF) (1:200) (Chemicon), and VEGFR-1 (1:150) (Epitomics, Inc.).

    Techniques: Isolation, FACS, In Vitro, Expressing

    MTUS1/ATIP3a overexpression is related to the migration and invasion of SACC. (A) Differential expression of ERK-Slug pathway in SACC-LM cells treated with either control vector or ATIP3a plasmid. SACC-LM cells displayed decreased Slug, Vimentin and pERK1/2 protein levels and increased E-cadherin, ATIP3a protein levels upon ATIP3a overexpression. ATIP3a protein was detected by anti-MTUS1 antibody. ( B and C ) The migration and invasion ability of SACC cells was assessed by a transwell migration and invasion assay. ATIP3a overexpression significantly inhibited the migration (B) and invasion (C) of SACC-LM cells. (D) Cell proliferation was measured using a MTT assay. The cell proliferation rate of SACC-LM was significantly inhibited after overexpressed with MTUS1/ATIP3a. *: P

    Journal: BMC Cancer

    Article Title: MTUS1/ATIP3a down-regulation is associated with enhanced migration, invasion and poor prognosis in salivary adenoid cystic carcinoma

    doi: 10.1186/s12885-015-1209-x

    Figure Lengend Snippet: MTUS1/ATIP3a overexpression is related to the migration and invasion of SACC. (A) Differential expression of ERK-Slug pathway in SACC-LM cells treated with either control vector or ATIP3a plasmid. SACC-LM cells displayed decreased Slug, Vimentin and pERK1/2 protein levels and increased E-cadherin, ATIP3a protein levels upon ATIP3a overexpression. ATIP3a protein was detected by anti-MTUS1 antibody. ( B and C ) The migration and invasion ability of SACC cells was assessed by a transwell migration and invasion assay. ATIP3a overexpression significantly inhibited the migration (B) and invasion (C) of SACC-LM cells. (D) Cell proliferation was measured using a MTT assay. The cell proliferation rate of SACC-LM was significantly inhibited after overexpressed with MTUS1/ATIP3a. *: P

    Article Snippet: Western blot analysis Western blots were performed as described previously [ ] using specific antibodies against E-cadtherin (E-cad), Vimentin (Vim), ERK1/2, p-ERK1/2, Slug, GFP, GAPDH (Cell Signaling Technology, Beverly, MA, USA), MTUS1 (Aviva, San Diego, CA, USA) and an Immu-Star HRP Substrate Kit (BIO-RAD, Hercules, CA, USA).

    Techniques: Over Expression, Migration, Expressing, Plasmid Preparation, Invasion Assay, MTT Assay

    MTUS1/ATIP3a knockdown promotes the migration and invasion of SACC. (A) Obviously reductions in ATIP3a protein levels were observed in the ATIP3a siRNA-transfected SACC-83 cells compared to the negative control-siRNA transfected cells. SACC-83 cells displayed increased Slug, Vimentin and pERK1/2 protein levels and decreased E-cadherin protein levels upon ATIP3a knockdown. (B, C) ATIP3a knockdown promotes the migration and invasion of SACC-83 cells. (D) The cell proliferation rate of the SACC-83 cells was significantly promoted after transfection with the ATIP3a siRNA. *: P

    Journal: BMC Cancer

    Article Title: MTUS1/ATIP3a down-regulation is associated with enhanced migration, invasion and poor prognosis in salivary adenoid cystic carcinoma

    doi: 10.1186/s12885-015-1209-x

    Figure Lengend Snippet: MTUS1/ATIP3a knockdown promotes the migration and invasion of SACC. (A) Obviously reductions in ATIP3a protein levels were observed in the ATIP3a siRNA-transfected SACC-83 cells compared to the negative control-siRNA transfected cells. SACC-83 cells displayed increased Slug, Vimentin and pERK1/2 protein levels and decreased E-cadherin protein levels upon ATIP3a knockdown. (B, C) ATIP3a knockdown promotes the migration and invasion of SACC-83 cells. (D) The cell proliferation rate of the SACC-83 cells was significantly promoted after transfection with the ATIP3a siRNA. *: P

    Article Snippet: Western blot analysis Western blots were performed as described previously [ ] using specific antibodies against E-cadtherin (E-cad), Vimentin (Vim), ERK1/2, p-ERK1/2, Slug, GFP, GAPDH (Cell Signaling Technology, Beverly, MA, USA), MTUS1 (Aviva, San Diego, CA, USA) and an Immu-Star HRP Substrate Kit (BIO-RAD, Hercules, CA, USA).

    Techniques: Migration, Transfection, Negative Control

    Effects of AGX51 on EMT signatures. Related to Figure 4. (A) Western blot for E-cadherin, Vimentin, Snail, Twist1 and Zeb1 on whole cell lysates from 4T1 cells treated with 40 µM AGX51 for 0-48, or 0-72 hours. Tubulin/Actin serve as protein loading controls.

    Journal: bioRxiv

    Article Title: Anti-tumor effects of an Id antagonist with no acquired resistance

    doi: 10.1101/2020.01.06.894840

    Figure Lengend Snippet: Effects of AGX51 on EMT signatures. Related to Figure 4. (A) Western blot for E-cadherin, Vimentin, Snail, Twist1 and Zeb1 on whole cell lysates from 4T1 cells treated with 40 µM AGX51 for 0-48, or 0-72 hours. Tubulin/Actin serve as protein loading controls.

    Article Snippet: Immunohistochemistry Slides mounted with sections from formalin-fixed and paraffin-embedded mouse tissues were deparaffinized, rehydrated and stained with hematoxylin and eosin (H & E), as well as with the following primary antibodies: Id1 (37-2, Biocheck 1:1000), Vimentin (5741, Cell Signaling 1:500), Cyclin D1 (2978, Cell Signaling 1:500).

    Techniques: Western Blot

    The influences of CAV3.1 knockdown on expressions of proteins involving cell cycle, EMT, and Akt activity in PCa cells. Notes: ( A ) CAV3.1 knockdown led to decreased CCND1, but not for CCNE1 and CCNB1 in PC-3 and DU145 cells. Moreover, CAV3.1 knockdown led to decreased N-cad and Vim, 2 mesenchymal markers, and increased E-cad, an epithelial marker. ( B ) CAV3.1 knockdown led to decreased p-Akt (S473) and p-Akt (T308) in PC-3 and DU145 cells. Abbreviations: E-cad, E-cadherin; EMT, epithelial–mesenchymal transition; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; N-cad, N-cadherin; PCa, prostate cancer.

    Journal: Cancer Management and Research

    Article Title: CAV3.1 knockdown suppresses cell proliferation, migration and invasion of prostate cancer cells by inhibiting AKT

    doi: 10.2147/CMAR.S172948

    Figure Lengend Snippet: The influences of CAV3.1 knockdown on expressions of proteins involving cell cycle, EMT, and Akt activity in PCa cells. Notes: ( A ) CAV3.1 knockdown led to decreased CCND1, but not for CCNE1 and CCNB1 in PC-3 and DU145 cells. Moreover, CAV3.1 knockdown led to decreased N-cad and Vim, 2 mesenchymal markers, and increased E-cad, an epithelial marker. ( B ) CAV3.1 knockdown led to decreased p-Akt (S473) and p-Akt (T308) in PC-3 and DU145 cells. Abbreviations: E-cad, E-cadherin; EMT, epithelial–mesenchymal transition; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; N-cad, N-cadherin; PCa, prostate cancer.

    Article Snippet: Subsequently, the membranes were subjected to blocking with 5% bovine serum albumin, incubation with the corresponding primary antibodies at 4°C overnight: CAV3.1(Santa Cruz Biotechnology), E-cadherin, N- cadherin, Vimentin (Vim), CCNB1, CCND1, CCNE1, and GAPDH (Abclonal, Woburn, MA, USA), p-Akt (S473), p-Akt (T308), and Akt (CST, Boston, MA, USA), and incubation with the secondary antibody for 1 hour at room temperature.

    Techniques: Activity Assay, Marker

    Ectopic expression of Akt reversed the inhibitory effects on proliferation and invasion of PCa cells caused by CAV3.1 knockdown. Notes: ( A ) Ectopic expression of Akt antagonized the effects of CAV3.1 knockdown on CCND1, E-cad, N-cad, Vim, p-Akt (S473), and p-Akt (T308) in PCa cells. Ectopic expression of Akt reversed the clone number ( B ), EdU incorporated rates ( C ), and invasive cells (magnification ×200) ( D ) in PC-3 and DU145 cells with CAV3 (magnification ×100).1 knockdown. * P

    Journal: Cancer Management and Research

    Article Title: CAV3.1 knockdown suppresses cell proliferation, migration and invasion of prostate cancer cells by inhibiting AKT

    doi: 10.2147/CMAR.S172948

    Figure Lengend Snippet: Ectopic expression of Akt reversed the inhibitory effects on proliferation and invasion of PCa cells caused by CAV3.1 knockdown. Notes: ( A ) Ectopic expression of Akt antagonized the effects of CAV3.1 knockdown on CCND1, E-cad, N-cad, Vim, p-Akt (S473), and p-Akt (T308) in PCa cells. Ectopic expression of Akt reversed the clone number ( B ), EdU incorporated rates ( C ), and invasive cells (magnification ×200) ( D ) in PC-3 and DU145 cells with CAV3 (magnification ×100).1 knockdown. * P

    Article Snippet: Subsequently, the membranes were subjected to blocking with 5% bovine serum albumin, incubation with the corresponding primary antibodies at 4°C overnight: CAV3.1(Santa Cruz Biotechnology), E-cadherin, N- cadherin, Vimentin (Vim), CCNB1, CCND1, CCNE1, and GAPDH (Abclonal, Woburn, MA, USA), p-Akt (S473), p-Akt (T308), and Akt (CST, Boston, MA, USA), and incubation with the secondary antibody for 1 hour at room temperature.

    Techniques: Expressing

    TAM-secreted CXCL1 is highly increased in the lung metastatic lesion of breast cancer. a TAMs were isolated from breast tumors by differential adhesion technique and validated by F4/80 + /CD206 + staining. b Cytokine array revealed that CXCL1 had the highest expression in the supernatants of TAMs. c Primary mammary tumors and lung metastatic lesions were collected from MMTV-PyVT +/- mice, respectively, and validated by HE staining. d CXCL1 expression was significantly enhanced in the lung metastatic lesions compared with its primary tumors, accompanied by increased expression levels of vimentin, N-cadherin and β-catenin, whereas the epithelial marker E-cadherin was reduced, indicating that CXCL1 expression was closely correlated with EMT process. Meanwhile, ARG1 expression was also increased in the lung metastasis lesions, implying that the enhanced CXCL1 expression might be correlated with increased M2 macrophage phenotype. (All values from three independent experiments are quantified as mean ± SD, * P

    Journal: Cell Death & Disease

    Article Title: CXCL1 derived from tumor-associated macrophages promotes breast cancer metastasis via activating NF-κB/SOX4 signaling

    doi: 10.1038/s41419-018-0876-3

    Figure Lengend Snippet: TAM-secreted CXCL1 is highly increased in the lung metastatic lesion of breast cancer. a TAMs were isolated from breast tumors by differential adhesion technique and validated by F4/80 + /CD206 + staining. b Cytokine array revealed that CXCL1 had the highest expression in the supernatants of TAMs. c Primary mammary tumors and lung metastatic lesions were collected from MMTV-PyVT +/- mice, respectively, and validated by HE staining. d CXCL1 expression was significantly enhanced in the lung metastatic lesions compared with its primary tumors, accompanied by increased expression levels of vimentin, N-cadherin and β-catenin, whereas the epithelial marker E-cadherin was reduced, indicating that CXCL1 expression was closely correlated with EMT process. Meanwhile, ARG1 expression was also increased in the lung metastasis lesions, implying that the enhanced CXCL1 expression might be correlated with increased M2 macrophage phenotype. (All values from three independent experiments are quantified as mean ± SD, * P

    Article Snippet: After blocking in 10% goat serum for 1 h, the cover slips were incubated with primary antibodies SOX4 (no. ab80261, ABclonal) or Vimentin (no. A2666, ABclonal) overnight at 4 °C.

    Techniques: Isolation, Staining, Expressing, Mouse Assay, Marker

    TAM-secreted CXCL1 promotes mouse breast cancer cells migration and invasion. a CXCL1 had little influence on the proliferation of breast cancer cells including 4T1, MDA-MB-231 and MCF-7 in both dose- and time-dependent manner. b Would-healing and Transwell assay revealed that CXCL1 could promote the migration and invasion ability of 4T1 cells. c Western blotting results showed that CXCL1 administration promotes the EMT process of 4T1 cells, presenting as dose-dependent increase of vimentin, N-cadherin, β-catenin and gradual downregulation of E-cadherin. Gelatin zymography also indicated that CXCL1 treatment results in increased secretion of MMP-9 and MMP-2 from 4T1 cells. d The mouse macrophage cell line Raw264.7 was induced to TAMs by administrating IL-4 and IL-13, resulting in the increased expression of CD206 and Arg-1, and reduction of iNOS; Meanwhile, ELISA assay demonstrated that CXCL1 expression in M2-Raw264.7 supernatants was significantly higher than that in either M0-Raw164.7 or 4T1 cancer cells. Notably, the CM of TAMs did not increase CXCL1 expression in 4T1 cancer cells. e TAMs-CM treatment led to increased migration and invasion ability of 4T1 cells, whereas was blocked by administration of CXCL1-neutralizing antibody. f, g CXCR2 silencing in 4T1 cancer cells did not block CXCL1-induced EMT and invasion ability. (All values from three independent experiments are quantified as mean ± SD, ** P

    Journal: Cell Death & Disease

    Article Title: CXCL1 derived from tumor-associated macrophages promotes breast cancer metastasis via activating NF-κB/SOX4 signaling

    doi: 10.1038/s41419-018-0876-3

    Figure Lengend Snippet: TAM-secreted CXCL1 promotes mouse breast cancer cells migration and invasion. a CXCL1 had little influence on the proliferation of breast cancer cells including 4T1, MDA-MB-231 and MCF-7 in both dose- and time-dependent manner. b Would-healing and Transwell assay revealed that CXCL1 could promote the migration and invasion ability of 4T1 cells. c Western blotting results showed that CXCL1 administration promotes the EMT process of 4T1 cells, presenting as dose-dependent increase of vimentin, N-cadherin, β-catenin and gradual downregulation of E-cadherin. Gelatin zymography also indicated that CXCL1 treatment results in increased secretion of MMP-9 and MMP-2 from 4T1 cells. d The mouse macrophage cell line Raw264.7 was induced to TAMs by administrating IL-4 and IL-13, resulting in the increased expression of CD206 and Arg-1, and reduction of iNOS; Meanwhile, ELISA assay demonstrated that CXCL1 expression in M2-Raw264.7 supernatants was significantly higher than that in either M0-Raw164.7 or 4T1 cancer cells. Notably, the CM of TAMs did not increase CXCL1 expression in 4T1 cancer cells. e TAMs-CM treatment led to increased migration and invasion ability of 4T1 cells, whereas was blocked by administration of CXCL1-neutralizing antibody. f, g CXCR2 silencing in 4T1 cancer cells did not block CXCL1-induced EMT and invasion ability. (All values from three independent experiments are quantified as mean ± SD, ** P

    Article Snippet: After blocking in 10% goat serum for 1 h, the cover slips were incubated with primary antibodies SOX4 (no. ab80261, ABclonal) or Vimentin (no. A2666, ABclonal) overnight at 4 °C.

    Techniques: Migration, Multiple Displacement Amplification, Transwell Assay, Western Blot, Zymography, Expressing, Enzyme-linked Immunosorbent Assay, Blocking Assay

    CXCL1 promotes breast cancer EMT process via activating SOX4 signaling. a qPCR screening assay revealed SOX4 as the highest responsive gene following CXCL1 administration in both MDA-MB-231 and MCF-7 cells. b Immunofluorescence assay revealed that CXCL1 administration resulted in increased SOX4 expression and its nucleus transportation ( × 400). c Western blotting assay indicated that CXCL1 overexpression resulted in SOX4 upregulation, accompanied by enhanced EMT process. By contrast, CXCL1 silencing in both breast cancer cells led to the downregulation of SOX4 and inhibited EMT process. d SOX4 silencing inhibited the EMT process in both breast cancer cells induced by CXCL1 administration, presenting as decreased expression of vimentin, β-catenin and upregulation of E-cadherin. (All values from three independent experiments are quantified as mean ± SD, ** P

    Journal: Cell Death & Disease

    Article Title: CXCL1 derived from tumor-associated macrophages promotes breast cancer metastasis via activating NF-κB/SOX4 signaling

    doi: 10.1038/s41419-018-0876-3

    Figure Lengend Snippet: CXCL1 promotes breast cancer EMT process via activating SOX4 signaling. a qPCR screening assay revealed SOX4 as the highest responsive gene following CXCL1 administration in both MDA-MB-231 and MCF-7 cells. b Immunofluorescence assay revealed that CXCL1 administration resulted in increased SOX4 expression and its nucleus transportation ( × 400). c Western blotting assay indicated that CXCL1 overexpression resulted in SOX4 upregulation, accompanied by enhanced EMT process. By contrast, CXCL1 silencing in both breast cancer cells led to the downregulation of SOX4 and inhibited EMT process. d SOX4 silencing inhibited the EMT process in both breast cancer cells induced by CXCL1 administration, presenting as decreased expression of vimentin, β-catenin and upregulation of E-cadherin. (All values from three independent experiments are quantified as mean ± SD, ** P

    Article Snippet: After blocking in 10% goat serum for 1 h, the cover slips were incubated with primary antibodies SOX4 (no. ab80261, ABclonal) or Vimentin (no. A2666, ABclonal) overnight at 4 °C.

    Techniques: Real-time Polymerase Chain Reaction, Screening Assay, Multiple Displacement Amplification, Immunofluorescence, Expressing, Western Blot, Over Expression

    TAM-secreted CXCL1 promotes human breast cancer cells migration and invasion. a Would-healing and Transwell assay revealed that CXCL1 could promote the migration and invasion ability of both MDA-MB-231 and MCF-7 cells. b Western blotting results showed that CXCL1 administration dose dependently promotes the EMT process of both breast cancer cells, presenting as increased expression of vimentin, N-cadherin, β-catenin and gradual downregulation of E-cadherin. c The enhanced expression levels of CD206, Arg1 and CXCL1 validated the successful induction of TAMs by administrating IL-4. Meanwhile, shCXCL1 was applied to knockdown its expression in M2-THP1 macrophages. d CXCL1 silencing in M2-THP1 macrophages suppressed the enhanced migration and invasion abilities of MDA-MB-231 and MCF-7 cells induced by TAMs-CM. (All values from three independent experiments are quantified as mean ± SD, ** P

    Journal: Cell Death & Disease

    Article Title: CXCL1 derived from tumor-associated macrophages promotes breast cancer metastasis via activating NF-κB/SOX4 signaling

    doi: 10.1038/s41419-018-0876-3

    Figure Lengend Snippet: TAM-secreted CXCL1 promotes human breast cancer cells migration and invasion. a Would-healing and Transwell assay revealed that CXCL1 could promote the migration and invasion ability of both MDA-MB-231 and MCF-7 cells. b Western blotting results showed that CXCL1 administration dose dependently promotes the EMT process of both breast cancer cells, presenting as increased expression of vimentin, N-cadherin, β-catenin and gradual downregulation of E-cadherin. c The enhanced expression levels of CD206, Arg1 and CXCL1 validated the successful induction of TAMs by administrating IL-4. Meanwhile, shCXCL1 was applied to knockdown its expression in M2-THP1 macrophages. d CXCL1 silencing in M2-THP1 macrophages suppressed the enhanced migration and invasion abilities of MDA-MB-231 and MCF-7 cells induced by TAMs-CM. (All values from three independent experiments are quantified as mean ± SD, ** P

    Article Snippet: After blocking in 10% goat serum for 1 h, the cover slips were incubated with primary antibodies SOX4 (no. ab80261, ABclonal) or Vimentin (no. A2666, ABclonal) overnight at 4 °C.

    Techniques: Migration, Transwell Assay, Multiple Displacement Amplification, Western Blot, Expressing

    HuCCT1 and Huh28 cell lines exhibit an epithelioid and a mesenchymal‐like phenotype respectively, the later correlating with endogenous activation of the TGFβ pathway. (A) Phase contrast micrographs of HuCCT1 and Huh28 cells (left panel; scale bar, 100 µm) and expression of vimentin and E‐cadherin at the RNA (middle panel) and protein level (right panel). (B) Volcano plot of genes differentially expressed between HuCCT1 and Huh28 (left panel) and GSEA highlighting a TGFβ signature enriched in the gene expression profile of Huh28. (C) Enzyme‐linked immunosorbent assay demonstrating increased secretion of TGFβ1 in the supernatant of Huh28 cells 24 hours after overnight serum starvation. (D) Relative luciferase activity of HuCCT1 and Huh28 engineered cell lines in which a luciferase gene is under the transcriptional control of a TATA box alone (white bars) or with SMAD responsive elements (black bars). Luciferase activity was measured at the basal level without exogenous addition of TGFβ after overnight serum starvation. (E) Gene expression analysis of TGFβ target genes in HuCCT1 and Huh28 cells at the basal level by microarray and Q‐RT‐PCR. (F) Q‐RT‐PCR analysis of TGFβ target gene SMAD7 in Huh28 cells treated with 10 µM TGFβ inhibitors (black bars). Statistical analysis was performed by t test (* P

    Journal: Hepatology Communications

    Article Title: A novel transforming growth factor beta‐induced long noncoding RNA promotes an inflammatory microenvironment in human intrahepatic cholangiocarcinoma

    doi: 10.1002/hep4.1142

    Figure Lengend Snippet: HuCCT1 and Huh28 cell lines exhibit an epithelioid and a mesenchymal‐like phenotype respectively, the later correlating with endogenous activation of the TGFβ pathway. (A) Phase contrast micrographs of HuCCT1 and Huh28 cells (left panel; scale bar, 100 µm) and expression of vimentin and E‐cadherin at the RNA (middle panel) and protein level (right panel). (B) Volcano plot of genes differentially expressed between HuCCT1 and Huh28 (left panel) and GSEA highlighting a TGFβ signature enriched in the gene expression profile of Huh28. (C) Enzyme‐linked immunosorbent assay demonstrating increased secretion of TGFβ1 in the supernatant of Huh28 cells 24 hours after overnight serum starvation. (D) Relative luciferase activity of HuCCT1 and Huh28 engineered cell lines in which a luciferase gene is under the transcriptional control of a TATA box alone (white bars) or with SMAD responsive elements (black bars). Luciferase activity was measured at the basal level without exogenous addition of TGFβ after overnight serum starvation. (E) Gene expression analysis of TGFβ target genes in HuCCT1 and Huh28 cells at the basal level by microarray and Q‐RT‐PCR. (F) Q‐RT‐PCR analysis of TGFβ target gene SMAD7 in Huh28 cells treated with 10 µM TGFβ inhibitors (black bars). Statistical analysis was performed by t test (* P

    Article Snippet: Human anti‐E‐cadherin (CDH1) (AF648; R & D Systems) and anti‐vimentin (VIM) (B01P; Abnova) antibodies were used.

    Techniques: Activation Assay, Expressing, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Microarray, Reverse Transcription Polymerase Chain Reaction

    BHLHE41 expression suppressed tumour cell migration and invasion of MCF‐7 cells and MDA‐MB‐231 cells. A, B, The mRNA levels of CLDN1, CLDN4 and CDH1 were up‐regulated, whereas those of SNAI1, SNAI2, VIM and CDH2 were down‐regulated after transfecting with exogenous BHLHE41 in MCF‐7 and MDA‐MB‐231 cells. C, D, WB analysis showed comparable results in the expression of the above genes. E‐H, Transwell assay for assessing cell migration and invasion of MCF‐7 and MDA‐MB‐231. As measured by the transwell assay without or with gel cover, BHLHE41 overexpression led to a significant decrease in the number of migrated and invasion cells. Transwell assays were repeated in triplicate. Ten views were selected for each trial, and the migrated or invasion cells were counted for the final statistical analysis. Each value represents the mean ± SE (bars) of three independent experiments, *** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: BHLHE41 suppresses MCF‐7 cell invasion via MAPK/JNK pathway. BHLHE41 suppresses MCF‐7 cell invasion via MAPK/JNK pathway

    doi: 10.1111/jcmm.15033

    Figure Lengend Snippet: BHLHE41 expression suppressed tumour cell migration and invasion of MCF‐7 cells and MDA‐MB‐231 cells. A, B, The mRNA levels of CLDN1, CLDN4 and CDH1 were up‐regulated, whereas those of SNAI1, SNAI2, VIM and CDH2 were down‐regulated after transfecting with exogenous BHLHE41 in MCF‐7 and MDA‐MB‐231 cells. C, D, WB analysis showed comparable results in the expression of the above genes. E‐H, Transwell assay for assessing cell migration and invasion of MCF‐7 and MDA‐MB‐231. As measured by the transwell assay without or with gel cover, BHLHE41 overexpression led to a significant decrease in the number of migrated and invasion cells. Transwell assays were repeated in triplicate. Ten views were selected for each trial, and the migrated or invasion cells were counted for the final statistical analysis. Each value represents the mean ± SE (bars) of three independent experiments, *** P

    Article Snippet: The membranes were incubated overnight at 4°C with the following primary antibodies: BHLHE41(1:500,Abcam), BHLHE40 (1:100, Sigma), GAPDH (1:2000, Sigma), His‐tag (1:1000, CWBio), Myc‐tag (1:1000, ImmunoWay), CLDN1/Claudin‐1 (1:500, Invitrogen), CLDN4/Claudin‐4 (1:500, Invitrogen), SNAI1/snail (1:500, Cell Signaling Technology), SNAI2/slug (1:500, Cell Signaling Technology), VIM/Vimentin (1:1000, BD Biosciences), CDH2/N‐cadherin (1:1000, BD Biosciences) and CDH1/E‐cadherin (1:500; ProteinTech Group, Inc).

    Techniques: Expressing, Migration, Multiple Displacement Amplification, Western Blot, Transwell Assay, Over Expression

    In vitro characterization of pituitary adenoma cell cultures for mesenchymal stromal cell markers. Representative FACS histograms of pituitary somatotroph adenoma cell culture (a–f). Cells were positive for CD90 (a), CD105 (b), and CD44 (c) and were negative for CD34 (d), CD45 (e), and HLA-DR (f). Representative immunostaining of VIM (green) (g) and negative control (h). Nuclei were stained with DAPI (blue). 200x magnification. CD90, Thy-1 cell surface antigen; CD105, endoglin; CD44, CD44 molecule (Indian blood group); CD34, CD34 molecule; CD45, protein tyrosine phosphatase, receptor type, C; HLA-DR, major histocompatibility complex, class II, DR.

    Journal: Stem Cells International

    Article Title: Functional Characteristics of Multipotent Mesenchymal Stromal Cells from Pituitary Adenomas

    doi: 10.1155/2016/7103720

    Figure Lengend Snippet: In vitro characterization of pituitary adenoma cell cultures for mesenchymal stromal cell markers. Representative FACS histograms of pituitary somatotroph adenoma cell culture (a–f). Cells were positive for CD90 (a), CD105 (b), and CD44 (c) and were negative for CD34 (d), CD45 (e), and HLA-DR (f). Representative immunostaining of VIM (green) (g) and negative control (h). Nuclei were stained with DAPI (blue). 200x magnification. CD90, Thy-1 cell surface antigen; CD105, endoglin; CD44, CD44 molecule (Indian blood group); CD34, CD34 molecule; CD45, protein tyrosine phosphatase, receptor type, C; HLA-DR, major histocompatibility complex, class II, DR.

    Article Snippet: Cells were incubated for 1 h with primary antibodies for vimentin (VIM), nestin (NES), glial fibrillary acidic protein (GFAP), ataxin 1 (ATXN1), tubulin, and beta 3 class III (TUBB3) (Santa Cruz Biotechnology, USA) as well as A2B5 (binds to neural tissue such as brain, spinal cord, and dorsal root ganglia), oligodendrocyte lineage transcription factor 2 (OLIG2), and solute carrier family 1 member 3 (SLC1A3) (Thermo Fisher Scientific, USA).

    Techniques: In Vitro, FACS, Cell Culture, Immunostaining, Negative Control, Staining

    Western blot showing APD-induced changes in the expression of the HIST1H4B, HSPA8, NCL, NPM1, and VIM proteins in C6 and B35 cells and the rat cortex. Nuclear protein extracts collected from APD-treated C6 cells, B35 cells and rat cortex were compared. a and ( b ), HIST1H4B; ( c ) and ( d ),HSPA8; ( e ) and ( f ), NCL; ( g ) and ( h ), NPM1; ( i ) and ( j ), VIM. Bar charts were constructed from triplicate western blot data obtained from three different batches of APD-treated cells or rat cortices using ANOVA followed by Dunnett’s test (* p -value

    Journal: BMC Pharmacology & Toxicology

    Article Title: Proteomic study revealed antipsychotics-induced nuclear protein regulations in B35 cells are similar to the regulations in C6 cells and rat cortex

    doi: 10.1186/s40360-018-0199-0

    Figure Lengend Snippet: Western blot showing APD-induced changes in the expression of the HIST1H4B, HSPA8, NCL, NPM1, and VIM proteins in C6 and B35 cells and the rat cortex. Nuclear protein extracts collected from APD-treated C6 cells, B35 cells and rat cortex were compared. a and ( b ), HIST1H4B; ( c ) and ( d ),HSPA8; ( e ) and ( f ), NCL; ( g ) and ( h ), NPM1; ( i ) and ( j ), VIM. Bar charts were constructed from triplicate western blot data obtained from three different batches of APD-treated cells or rat cortices using ANOVA followed by Dunnett’s test (* p -value

    Article Snippet: Antibodies The antibodies included: anti-actin, ab6276 (Abcam, Cambridge, UK); anti-Histone H4 (HIST1H4B), #13919 (Cell Signaling Technology, Danvers, MA, USA); anti- Heat shock cognate 71 kDa protein (HSPA8), #8444 (Cell Signaling Technology); anti-nucleolin (NCL), #12247 (Cell Signaling Technology); anti-nucleophosmin (NPM1), #3542 (Cell Signaling Technology); anti-vimentin (VIM), #3932 (Cell Signaling Technology); anti-plectin-1 (PLEC), #12254 (Cell Signaling Technology).

    Techniques: Western Blot, Expressing, Construct

    TGF-β1 induced EMT progress and restored migration, invasion abilities suppressed by Neferine. (a,b) TGF-β1 induced EMT progress. HCC cells were treated with 5 ng/ml, 10 ng/ml, 20 ng/ml TGF-β1 for 24 hrs or 48 hrs to induce EMT. mRNA and protein expression of EMT biomarkers (E-cadherin, N-cadherin Vimentin), and EMT promoting transcription factor (Snail) was determined by qRT-PCR and by Western blot. Original blots of high-contrast blots are presented in Supplementary Fig. S4 . (c) Morphological changes of HCC cells treated with 10 ng/ml TGF-β1 for different times. TGF-β1 changed cells morphology from pebble-like epithelial to dispersed, spindle-like mesenchymal and pseudopodium stretching. (Original magnification: ×400).

    Journal: Scientific Reports

    Article Title: The anti-tumor activities of Neferine on cell invasion and oxaliplatin sensitivity regulated by EMT via Snail signaling in hepatocellular carcinoma

    doi: 10.1038/srep41616

    Figure Lengend Snippet: TGF-β1 induced EMT progress and restored migration, invasion abilities suppressed by Neferine. (a,b) TGF-β1 induced EMT progress. HCC cells were treated with 5 ng/ml, 10 ng/ml, 20 ng/ml TGF-β1 for 24 hrs or 48 hrs to induce EMT. mRNA and protein expression of EMT biomarkers (E-cadherin, N-cadherin Vimentin), and EMT promoting transcription factor (Snail) was determined by qRT-PCR and by Western blot. Original blots of high-contrast blots are presented in Supplementary Fig. S4 . (c) Morphological changes of HCC cells treated with 10 ng/ml TGF-β1 for different times. TGF-β1 changed cells morphology from pebble-like epithelial to dispersed, spindle-like mesenchymal and pseudopodium stretching. (Original magnification: ×400).

    Article Snippet: 5 μm-thick sections were then stained against Ki67 (1:200, Abcam), E-cadherin (1:200, Abcam) and Vimentin (1:400, Abcam).

    Techniques: Migration, Expressing, Quantitative RT-PCR, Western Blot

    Knockdown of Snail inhibited EMT progress, migration/invasion capacities and enhanced OXA sensitivity. (a) qRT-PCR and Western blot confirmed decreased expression level of Snail in si-Snail groups. Knockdown of Snail caused the changes of EMT biomarkers with E-cadherin upregulation and downregulation of N-cadherin/Vimentin. (b,c) Knockdown of Snail inhibited migration and invasion capacities of HCC in wound healing assays and transwell invasion assays. si-control vs. si-Snail, p

    Journal: Scientific Reports

    Article Title: The anti-tumor activities of Neferine on cell invasion and oxaliplatin sensitivity regulated by EMT via Snail signaling in hepatocellular carcinoma

    doi: 10.1038/srep41616

    Figure Lengend Snippet: Knockdown of Snail inhibited EMT progress, migration/invasion capacities and enhanced OXA sensitivity. (a) qRT-PCR and Western blot confirmed decreased expression level of Snail in si-Snail groups. Knockdown of Snail caused the changes of EMT biomarkers with E-cadherin upregulation and downregulation of N-cadherin/Vimentin. (b,c) Knockdown of Snail inhibited migration and invasion capacities of HCC in wound healing assays and transwell invasion assays. si-control vs. si-Snail, p

    Article Snippet: 5 μm-thick sections were then stained against Ki67 (1:200, Abcam), E-cadherin (1:200, Abcam) and Vimentin (1:400, Abcam).

    Techniques: Migration, Quantitative RT-PCR, Western Blot, Expressing

    The reversion effects of Neferine on TGF-β1-induced EMT model in HCC cells. (a,b) HCC cells were firstly treated with TGF-β1 for 48 hrs to induce EMT before the administration of Neferine (48 hrs) and/or OXA (48 hrs). mRNA and protein expression of epithelial marker (E-cadherin), mesenchymal markers (N-cadherin Vimentin), and EMT promoting transcription factor (Snail, Slug, Twist, Zeb1), which was determined by qRT-PCR and by Western blot in TGF-β1-treated HCC cells treated with Neferine and/or OXA. Original blots of high-contrast blots are presented in Supplementary Fig. S5 . (c) Representative double immunofluorescence staining for expression and co-localization of E-cadherin and Vimentin in TGF-β1-treated HCC cells treated with Neferine and/or OXA. (Original magnification: ×400). (d) HCC cells treated with 10 ng/ml TGF-β1 (TGF-β1 groups) or co-treated with 10 ng/ml TGF-β1 and Neferine (TGF-β1 + Neferine groups: applied Neferine to TGF-β1-treated HCC cells for 48 hrs). In Neferine + TGF-β1 groups, HCC cells were pre-treated with Neferine for 48 hrs before the administration of TGF-β1). Migration abilities of HepG2 and Bel-7402 cells were inhibited by Neferine in vitro and were canceled by TGF-β1 in wound healing assays. #: TGF-β1 vs. control, p

    Journal: Scientific Reports

    Article Title: The anti-tumor activities of Neferine on cell invasion and oxaliplatin sensitivity regulated by EMT via Snail signaling in hepatocellular carcinoma

    doi: 10.1038/srep41616

    Figure Lengend Snippet: The reversion effects of Neferine on TGF-β1-induced EMT model in HCC cells. (a,b) HCC cells were firstly treated with TGF-β1 for 48 hrs to induce EMT before the administration of Neferine (48 hrs) and/or OXA (48 hrs). mRNA and protein expression of epithelial marker (E-cadherin), mesenchymal markers (N-cadherin Vimentin), and EMT promoting transcription factor (Snail, Slug, Twist, Zeb1), which was determined by qRT-PCR and by Western blot in TGF-β1-treated HCC cells treated with Neferine and/or OXA. Original blots of high-contrast blots are presented in Supplementary Fig. S5 . (c) Representative double immunofluorescence staining for expression and co-localization of E-cadherin and Vimentin in TGF-β1-treated HCC cells treated with Neferine and/or OXA. (Original magnification: ×400). (d) HCC cells treated with 10 ng/ml TGF-β1 (TGF-β1 groups) or co-treated with 10 ng/ml TGF-β1 and Neferine (TGF-β1 + Neferine groups: applied Neferine to TGF-β1-treated HCC cells for 48 hrs). In Neferine + TGF-β1 groups, HCC cells were pre-treated with Neferine for 48 hrs before the administration of TGF-β1). Migration abilities of HepG2 and Bel-7402 cells were inhibited by Neferine in vitro and were canceled by TGF-β1 in wound healing assays. #: TGF-β1 vs. control, p

    Article Snippet: 5 μm-thick sections were then stained against Ki67 (1:200, Abcam), E-cadherin (1:200, Abcam) and Vimentin (1:400, Abcam).

    Techniques: Expressing, Marker, Quantitative RT-PCR, Western Blot, Double Immunofluorescence Staining, Migration, In Vitro

    The xenograft tumors displayed characteristic histological features of the original anaplastic EPN, describing the following tests: Immunohistochemical xenograft tumors (EPN1-5) analysis with Prussian Blue for MION-Rh detection, immunohistochemical assay of tumor for glial fibrillary acidic protein (GFAP), hematoxylin and eosin staining, immunohistochemical assay of tumor for Ki67 to detect cell proliferation, vimentin (VIM) to intermediate filament, Nestin to neuron precursor and synaptophysin (SYP) to neuron mature. Black circle: arrangement pseudorosettes with classic intracellular lumens; black arrow: EPN cells that proliferate in the ependymal channel light; white arrow: EPN cells that invade in surrounding region; yellow arrow: hemosiderin granules.

    Journal: Oncotarget

    Article Title: Establishment of primary cell culture and an intracranial xenograft model of pediatric ependymoma: a prospect for therapy development and understanding of tumor biology

    doi: 10.18632/oncotarget.24932

    Figure Lengend Snippet: The xenograft tumors displayed characteristic histological features of the original anaplastic EPN, describing the following tests: Immunohistochemical xenograft tumors (EPN1-5) analysis with Prussian Blue for MION-Rh detection, immunohistochemical assay of tumor for glial fibrillary acidic protein (GFAP), hematoxylin and eosin staining, immunohistochemical assay of tumor for Ki67 to detect cell proliferation, vimentin (VIM) to intermediate filament, Nestin to neuron precursor and synaptophysin (SYP) to neuron mature. Black circle: arrangement pseudorosettes with classic intracellular lumens; black arrow: EPN cells that proliferate in the ependymal channel light; white arrow: EPN cells that invade in surrounding region; yellow arrow: hemosiderin granules.

    Article Snippet: Coronal sections were cut to 40 μm in thickness using a cryostat (Leica) and stained using standard procedures for hematoxylin-eosin and Prussian Blue staining and for IHC staining we used human primary antibodies included the following: antisynaptophysin (anti-SYP) (1:200), glial fibrillary acidic protein (GFAP) (1:200), vimentin (VIM) (1:200), epithelial membrane antigen (EMA) (1:100), MAP-2 (1:200), Ki-67 (1:20), p53 (1:100) (Abcam, Inc.), anti-Nestin (NES) (1:500) and VEGFR-1 (1:150) (Epitomics, Inc.).

    Techniques: Immunohistochemistry, Staining

    GATA3 upregulates E-cadherin and downregulates the expression of N-cadherin and vimentin in OS cells. Notes: ( A , B ) GATA3 was overexpressed in U2OS and Saos2 cells. The expression of EMT markers, such as epithelial marker (E-cadherin) and mesenchymal markers (N-cadherin and vimentin), were detected by qRT-PCR and western blotting assay. *

    Journal: OncoTargets and therapy

    Article Title: GATA3 is downregulated in osteosarcoma and facilitates EMT as well as migration through regulation of slug

    doi: 10.2147/OTT.S176534

    Figure Lengend Snippet: GATA3 upregulates E-cadherin and downregulates the expression of N-cadherin and vimentin in OS cells. Notes: ( A , B ) GATA3 was overexpressed in U2OS and Saos2 cells. The expression of EMT markers, such as epithelial marker (E-cadherin) and mesenchymal markers (N-cadherin and vimentin), were detected by qRT-PCR and western blotting assay. *

    Article Snippet: Subsequently, the membranes were incubated with antibodies, such as GATA3 (1:2,000, Abcam, Cambridge, UK), E-cadherin antibody (1:2,000, Abcam), N-cadherin antibody (1:2,000, Abcam), vimentin antibody (1:2,000, Abcam), slug (1:1,000, Abcam), snail (1:2,000, Abcam), twist1 (1:1,000, Abcam), and β-actin antibody (1:5,000, Abcam) overnight at 4°C.

    Techniques: Expressing, Marker, Quantitative RT-PCR, Western Blot

    Melatonin reverses 17β-estradiol-induced EMT in EEC involving Notch1 signaling pathway. EEC were treated with different drugs for 72 h. a The protein expression levels of Notch1 (NICD), E-cadherin, N-cadherin, Vimentin, Slug, Snail, Numb and Actin were determined by western blot. β-actin was used as a loading control. b The ratios of Notch1/E-cadherin/N-cadherin/Vimentin/Slug/Snail/Numb to β-actin were analyzed. Data are presented as the mean ± SD. E2: 17β-estradiol; MLT: melatonin. * p

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Melatonin inhibits 17β-estradiol-induced migration, invasion and epithelial-mesenchymal transition in normal and endometriotic endometrial epithelial cells

    doi: 10.1186/s12958-018-0375-5

    Figure Lengend Snippet: Melatonin reverses 17β-estradiol-induced EMT in EEC involving Notch1 signaling pathway. EEC were treated with different drugs for 72 h. a The protein expression levels of Notch1 (NICD), E-cadherin, N-cadherin, Vimentin, Slug, Snail, Numb and Actin were determined by western blot. β-actin was used as a loading control. b The ratios of Notch1/E-cadherin/N-cadherin/Vimentin/Slug/Snail/Numb to β-actin were analyzed. Data are presented as the mean ± SD. E2: 17β-estradiol; MLT: melatonin. * p

    Article Snippet: Rabbit anti-human E-Cadherin, N-Cadherin and Vimentin primary antibodies were obtained from Abcam (Cambridge, MA, USA).

    Techniques: Expressing, Western Blot

    The expression of EMT and Notch signaling pathway-related markers in NEC. NEC were treated with different drugs for 72 h. a The protein expression of Notch1 (NICD), E-cadherin, N-cadherin, Vimentin, Slug, Snail, Numb and Actin were determined by western blot. β-actin was used as a loading control. b The ratios of Notch1/ E-cadherin/ N-cadherin/Vimentin/Slug/ Snail/Numb to β-actin were analyzed. Data are presented as the mean ± SD. E2: 17β-estradiol; MLT: melatonin. * p

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Melatonin inhibits 17β-estradiol-induced migration, invasion and epithelial-mesenchymal transition in normal and endometriotic endometrial epithelial cells

    doi: 10.1186/s12958-018-0375-5

    Figure Lengend Snippet: The expression of EMT and Notch signaling pathway-related markers in NEC. NEC were treated with different drugs for 72 h. a The protein expression of Notch1 (NICD), E-cadherin, N-cadherin, Vimentin, Slug, Snail, Numb and Actin were determined by western blot. β-actin was used as a loading control. b The ratios of Notch1/ E-cadherin/ N-cadherin/Vimentin/Slug/ Snail/Numb to β-actin were analyzed. Data are presented as the mean ± SD. E2: 17β-estradiol; MLT: melatonin. * p

    Article Snippet: Rabbit anti-human E-Cadherin, N-Cadherin and Vimentin primary antibodies were obtained from Abcam (Cambridge, MA, USA).

    Techniques: Expressing, Western Blot

    Development and analysis of the establishment of a Stupp-like protocol in mice. A) At sacrifice, brains were collected, fixed in PFA, embedded in paraffin prior to be analyzed using immunohistochemistry using anti-vimentin antibodies (CTR: no injection; NS: no surgery; S: surgery only; SIT:surgery/irradiation/TMZ). B) Kaplan-Meier representation of mouse survival under this regiment (green: non-resected, blue: surgical resection, orange: surgical resection + radio/chemotherapy). C-D) Immunohistochemical analysis and quantitation of tumor angiogenesis post-resection. E-F) Immunohistochemical analysis and quantitation of tumor infiltration/invasion post-resection.

    Journal: bioRxiv

    Article Title: Local intracerebral Inhibition of IRE1 by MKC8866 sensitizes glioblastoma to irradiation/chemotherapy in vivo

    doi: 10.1101/841296

    Figure Lengend Snippet: Development and analysis of the establishment of a Stupp-like protocol in mice. A) At sacrifice, brains were collected, fixed in PFA, embedded in paraffin prior to be analyzed using immunohistochemistry using anti-vimentin antibodies (CTR: no injection; NS: no surgery; S: surgery only; SIT:surgery/irradiation/TMZ). B) Kaplan-Meier representation of mouse survival under this regiment (green: non-resected, blue: surgical resection, orange: surgical resection + radio/chemotherapy). C-D) Immunohistochemical analysis and quantitation of tumor angiogenesis post-resection. E-F) Immunohistochemical analysis and quantitation of tumor infiltration/invasion post-resection.

    Article Snippet: For immunohistochemistry staining, primary antibodies against vimentin (rabbit monoclonal, used at 1 in 250 dilution) was obtained from Abcam (Paris, France ); CD31 (rabbit polyclonal, used at 1 in 50 dilution) from Bioss Inc. (Cliniscience, Nanterre, France ); and against IBA-1 (rabbit polyclonal, used at 1 in 50 dilution) from Wako (Sobioda, Montbonnot, France ).

    Techniques: Mouse Assay, Immunohistochemistry, Injection, Irradiation, Quantitation Assay

    Soft substrate induces a rapid and homogeneous epithelial differentiation of RLSCs. (a) Phase-contrast micrographs of RLSCs grown on Petri plastic dish (CTRL;  E >  1 GPa) and on hydrogels with  E  = 0.4 kPa and 80 kPa, for 24 and 48 hours. Images are representative of three independent experiments. Scale bar: 100  μ m. (b) Phase-contrast micrographs and immunofluorescence of cells cultured on plastic (CTRL), 0.4 kPa and 80 kPa for 24 and 48 hours, stained for HNF4 α , Vimentin, and E-cadherin. The nuclei were stained with DAPI. Images are representative of three independent experiments. Scale bar: 50  μ m. (c) Western blot analysis of Vimentin at 24 and 48 hours after seeding on substrates with the indicated  E  values. CDK4 was used as a loading control.

    Journal: Stem Cells International

    Article Title: Modulating the Substrate Stiffness to Manipulate Differentiation of Resident Liver Stem Cells and to Improve the Differentiation State of Hepatocytes

    doi: 10.1155/2016/5481493

    Figure Lengend Snippet: Soft substrate induces a rapid and homogeneous epithelial differentiation of RLSCs. (a) Phase-contrast micrographs of RLSCs grown on Petri plastic dish (CTRL; E > 1 GPa) and on hydrogels with E = 0.4 kPa and 80 kPa, for 24 and 48 hours. Images are representative of three independent experiments. Scale bar: 100  μ m. (b) Phase-contrast micrographs and immunofluorescence of cells cultured on plastic (CTRL), 0.4 kPa and 80 kPa for 24 and 48 hours, stained for HNF4 α , Vimentin, and E-cadherin. The nuclei were stained with DAPI. Images are representative of three independent experiments. Scale bar: 50  μ m. (c) Western blot analysis of Vimentin at 24 and 48 hours after seeding on substrates with the indicated E values. CDK4 was used as a loading control.

    Article Snippet: Blots were blocked for 1 hour in 5% nonfat milk (Bio-Rad Laboratories, Inc., Hercules, CA, USA) prepared in TBST 1x and incubated overnight with the following primary antibodies: α -ERK1 (K23; 1 : 1000) and α -CDK4 (c-22; 1 : 1000) (Santa Cruz Biotechnology, Inc., CA, USA), phospho-p44/42 MAPK ERK1/2 (4370; 1 : 1000) (Cell Signalling Technology Boston, USA), and α -Vimentin (Clone V9; 1 : 1000) (Millipore, MA, USA).

    Techniques: Immunofluorescence, Cell Culture, Staining, Western Blot

    PBMMPs needs lymphocytes' support, and expresses both mesenchymal andhematopoietic features. (A) After 7 days, stromal-like cells took the most parts of adherent cells (a,b). Clone formation in the ensuing 5∼10 days cells (c). When floating cells (presumably T lymphocytes) were washed away as possible, cell proliferation ceased and karyopyknosis and fragmentation occurred (d). (B) Col-I/III genes expression under the control of β-actin was shown by RT-PCR. (C) PBMMP was positive for mesenchymal stem cell markers BMPR-IA, BMPR-II and Endoglin (CD105), the cytoskeletal component Vim, the fibroblast products Col-I, Col-II and Fn, the myofibroblast marker α-SMA and hematopoietic stem cell marker CD34 by immunofluorescence and immunohistochemistry. (Magnification 100×)

    Journal: International Journal of Medical Sciences

    Article Title: A Novel Population of Mesenchymal Progenitors with Hematopoietic Potential Originated from CD14- Peripheral Blood Mononuclear Cells

    doi:

    Figure Lengend Snippet: PBMMPs needs lymphocytes' support, and expresses both mesenchymal andhematopoietic features. (A) After 7 days, stromal-like cells took the most parts of adherent cells (a,b). Clone formation in the ensuing 5∼10 days cells (c). When floating cells (presumably T lymphocytes) were washed away as possible, cell proliferation ceased and karyopyknosis and fragmentation occurred (d). (B) Col-I/III genes expression under the control of β-actin was shown by RT-PCR. (C) PBMMP was positive for mesenchymal stem cell markers BMPR-IA, BMPR-II and Endoglin (CD105), the cytoskeletal component Vim, the fibroblast products Col-I, Col-II and Fn, the myofibroblast marker α-SMA and hematopoietic stem cell marker CD34 by immunofluorescence and immunohistochemistry. (Magnification 100×)

    Article Snippet: Cells were then incubated with one of the following mouse mAbs: anti-Fibronectin (Fn) (0.1 µg/ml), anti-Vimentin (Vim) (5 µg/ml; Chemicon, USA) and α-smooth muscle actin (SMA) (1:100; Santa cruz) overnight at 4℃ followed by treatment with second antibody.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, IA, Marker, Immunofluorescence, Immunohistochemistry

    VGLUT2-IR terminals were located in close apposition to tanycytes in the median eminence. A : vimentin-IR tanycyte processes (pseudo-colored blue) extend into the external zone of the median eminence in close proximity to CRH fibers (pseudo-colored red) and VGLUT2-IR (pseudo-colored green). Scale bar = 100 µm. B : higher-magnification image of dashed box in A . Arrows indicate VGLUT2 terminals surrounding tanycyte processes. An asterisk denotes close association of CRH fiber (pCRH-AAV-eGFP from PVN) with VGLUT2-IR. Scale bar = 25 µm.

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Acute hypoxia activates neuroendocrine, but not presympathetic, neurons in the paraventricular nucleus of the hypothalamus: differential role of nitric oxide

    doi: 10.1152/ajpregu.00543.2016

    Figure Lengend Snippet: VGLUT2-IR terminals were located in close apposition to tanycytes in the median eminence. A : vimentin-IR tanycyte processes (pseudo-colored blue) extend into the external zone of the median eminence in close proximity to CRH fibers (pseudo-colored red) and VGLUT2-IR (pseudo-colored green). Scale bar = 100 µm. B : higher-magnification image of dashed box in A . Arrows indicate VGLUT2 terminals surrounding tanycyte processes. An asterisk denotes close association of CRH fiber (pCRH-AAV-eGFP from PVN) with VGLUT2-IR. Scale bar = 25 µm.

    Article Snippet: No eGFP fluorescence was visualized in the PVN 4 wk after injection of a control empty (null) vector ( n = 3; example , B1 and B2 ; camera settings equivalent to , C1 and C2 ). eGFP-expressing fibers in the median eminence were identified in rats in which CRH neurons in the PVN had been transfected by prior injection of pCRH-AAV-eGFP. eGFP expression was evaluated in combination with CRH-IR ( n = 5) or in combination with vimentin-IR (marker for tanycytes; mouse anti-vimentin; 1:2,000; EMD Millipore, Temecula, CA) and VGLUT2-IR (guinea pig anti-VGLUT2; 1:1,000; EMD Millipore) ( n = 4).

    Techniques:

    PC3 cells are less tumorigenic and invasive after knockdown of PTK6 A) E-cadherin is increased upon knockdown of PTK6 in PC3 cells. PC3 cells were transfected with PTK6 siRNAs or control siRNAs for 2, 4 or 6 days. Total cell lysates were analyzed by immunoblotting with anti-E-cadherin, ZEB1, PTK6 and β-actin antibodies. Relative levels of E-cadherin, vimentin, and ZEB1 expression normalized to actin levels are indicated below the blots. B) A growth curve of PC3 cells transfected with PTK6 siRNAs or control siRNAs shows decreased proliferation from day 1 to day 7 after PTK6 knockdown. Relative Light Units (RLU) were measured by CellTiter-Glo Luminescent Cell Viability Assay. C) The number of colonies that form on plates 2 weeks post-plating is decreased upon PTK6 knockdown. Corresponding images are shown below the graph. D) The number of colonies that form in soft agar 3 weeks post-plating is decreased upon PTK6 knockdown. Representative images are shown. E) Cell invasion is impaired upon PTK6 knockdown in Matrigel invasion chamber assays. F) Knockdown of PTK6 in PC3 cells largely reduces metastases in SCID mice. PC3 cells stably expressing luciferase were transfected with PTK6 siRNA or control siRNA twice before injection. Mice were monitored under IVIS spectrum imaging system every week.

    Journal: Cancer research

    Article Title: PTK6 activation at the membrane regulates epithelial-mesenchymal transition in prostate cancer

    doi: 10.1158/0008-5472.CAN-13-0443

    Figure Lengend Snippet: PC3 cells are less tumorigenic and invasive after knockdown of PTK6 A) E-cadherin is increased upon knockdown of PTK6 in PC3 cells. PC3 cells were transfected with PTK6 siRNAs or control siRNAs for 2, 4 or 6 days. Total cell lysates were analyzed by immunoblotting with anti-E-cadherin, ZEB1, PTK6 and β-actin antibodies. Relative levels of E-cadherin, vimentin, and ZEB1 expression normalized to actin levels are indicated below the blots. B) A growth curve of PC3 cells transfected with PTK6 siRNAs or control siRNAs shows decreased proliferation from day 1 to day 7 after PTK6 knockdown. Relative Light Units (RLU) were measured by CellTiter-Glo Luminescent Cell Viability Assay. C) The number of colonies that form on plates 2 weeks post-plating is decreased upon PTK6 knockdown. Corresponding images are shown below the graph. D) The number of colonies that form in soft agar 3 weeks post-plating is decreased upon PTK6 knockdown. Representative images are shown. E) Cell invasion is impaired upon PTK6 knockdown in Matrigel invasion chamber assays. F) Knockdown of PTK6 in PC3 cells largely reduces metastases in SCID mice. PC3 cells stably expressing luciferase were transfected with PTK6 siRNA or control siRNA twice before injection. Mice were monitored under IVIS spectrum imaging system every week.

    Article Snippet: Anti-β-actin (AC-15) and vimentin antibodies were purchased from Sigma-Aldrich.

    Techniques: Transfection, Expressing, Cell Viability Assay, Mouse Assay, Stable Transfection, Luciferase, Injection, Imaging

    Active PTK6 at the plasma membrane promotes EMT in prostate tumor cells A) ) and it was used as a loading control. B) A subcellular fractionation assay was performed using PC3 cells expressing Palm-PTK6-YF or vector, and immunoblot analysis was performed using anti-E-cadherin, vimentin, ZEB1, P-PTK6 (PY342) and PTK6 antibodies. Both short and long exposures of PTK6 immunoblot are shown. C) An uncropped blot is presented to show specificity of the PY342 antibody. An arrowhead points to endogenous active PTK6 localized at the membrane in PC3 cells (Vector). Ectopic Palm-PTK6-YF runs slightly above the endogenous band in transfected cells (Paml-YF). D) mRNA levels of EMT markers are deregulated in Palm-PTK6-YF expressing PC3 cells. Quantitative RT-PCR was performed, and E-cadherin, vimentin, SLUG, Twist and ZEB1 mRNA levels were normalized to cyclophilin mRNA levels. *, P

    Journal: Cancer research

    Article Title: PTK6 activation at the membrane regulates epithelial-mesenchymal transition in prostate cancer

    doi: 10.1158/0008-5472.CAN-13-0443

    Figure Lengend Snippet: Active PTK6 at the plasma membrane promotes EMT in prostate tumor cells A) ) and it was used as a loading control. B) A subcellular fractionation assay was performed using PC3 cells expressing Palm-PTK6-YF or vector, and immunoblot analysis was performed using anti-E-cadherin, vimentin, ZEB1, P-PTK6 (PY342) and PTK6 antibodies. Both short and long exposures of PTK6 immunoblot are shown. C) An uncropped blot is presented to show specificity of the PY342 antibody. An arrowhead points to endogenous active PTK6 localized at the membrane in PC3 cells (Vector). Ectopic Palm-PTK6-YF runs slightly above the endogenous band in transfected cells (Paml-YF). D) mRNA levels of EMT markers are deregulated in Palm-PTK6-YF expressing PC3 cells. Quantitative RT-PCR was performed, and E-cadherin, vimentin, SLUG, Twist and ZEB1 mRNA levels were normalized to cyclophilin mRNA levels. *, P

    Article Snippet: Anti-β-actin (AC-15) and vimentin antibodies were purchased from Sigma-Aldrich.

    Techniques: Fractionation, Expressing, Plasmid Preparation, Transfection, Quantitative RT-PCR

    PTK6 mediated EMT occurs partially through increased AKT activity A) Increased AKT signaling in PC3 cells expressing Palm-PTK6-YF. Immunoblot analysis of total cell lysates of PC3 cells stably expressing Palm-PTK6-YF or vector was performed using anti-AKT, P-AKT (Thr308), P-AKT (Ser473), P-GSK3β (Ser9), SLUG PTK6 and β-catenin antibodies. Relative levels of P-AKT, P-GSK3β and SLUG normalized are indicated below the blots. B) Increased nuclear localization of SLUG in PC3 cells stably expressing Palm-PTK6-YF. Cells were stained with anti-SLUG antibody and counterstained with DAPI (blue). Size bar denotes 50 μm. C) Knockdown of AKT partially rescues Palm-PTK6-YF-induced EMT. PC3 cells expressing Palm-PTK6-YF or vector were transfected with AKT siRNAs or control siRNAs for 3 days. Immunoblotting was performed with anti-AKT, E-cadherin, vimentin, PTK6 and β-catenin antibodies. Relative levels of E-cadherin and vimentin normalized to the β-catenin loading control are indicated below the blots. D) Knockdown of p130CAS partially rescues Palm-PTK6-YF-induced EMT. PC3 cells expressing Palm-PTK6-YF or vector were transfected with p130CAS siRNAs or control siRNAs for 3 days. Immunoblotting was performed with anti-p130CAS, AKT, P-AKT (Thr308), P-GSK3β (Ser9), E-cadherin, vimentin, Myc-tag and β-catenin antibodies.

    Journal: Cancer research

    Article Title: PTK6 activation at the membrane regulates epithelial-mesenchymal transition in prostate cancer

    doi: 10.1158/0008-5472.CAN-13-0443

    Figure Lengend Snippet: PTK6 mediated EMT occurs partially through increased AKT activity A) Increased AKT signaling in PC3 cells expressing Palm-PTK6-YF. Immunoblot analysis of total cell lysates of PC3 cells stably expressing Palm-PTK6-YF or vector was performed using anti-AKT, P-AKT (Thr308), P-AKT (Ser473), P-GSK3β (Ser9), SLUG PTK6 and β-catenin antibodies. Relative levels of P-AKT, P-GSK3β and SLUG normalized are indicated below the blots. B) Increased nuclear localization of SLUG in PC3 cells stably expressing Palm-PTK6-YF. Cells were stained with anti-SLUG antibody and counterstained with DAPI (blue). Size bar denotes 50 μm. C) Knockdown of AKT partially rescues Palm-PTK6-YF-induced EMT. PC3 cells expressing Palm-PTK6-YF or vector were transfected with AKT siRNAs or control siRNAs for 3 days. Immunoblotting was performed with anti-AKT, E-cadherin, vimentin, PTK6 and β-catenin antibodies. Relative levels of E-cadherin and vimentin normalized to the β-catenin loading control are indicated below the blots. D) Knockdown of p130CAS partially rescues Palm-PTK6-YF-induced EMT. PC3 cells expressing Palm-PTK6-YF or vector were transfected with p130CAS siRNAs or control siRNAs for 3 days. Immunoblotting was performed with anti-p130CAS, AKT, P-AKT (Thr308), P-GSK3β (Ser9), E-cadherin, vimentin, Myc-tag and β-catenin antibodies.

    Article Snippet: Anti-β-actin (AC-15) and vimentin antibodies were purchased from Sigma-Aldrich.

    Techniques: Activity Assay, Expressing, Stable Transfection, Plasmid Preparation, Staining, Transfection

    Aberrant activation of PTK6 is accompanied by deregulated E-cadherin at the plasma membrane in prostate tumor cells of PB-Cre4, Pten flox/flox mice A) An enlarged anterior prostate was observed in PB-Cre4, Pten flox/flox mice at the age of 6 months. B) Endogenous PTK6 is activated at the membrane in prostate tumor cells in a murine model (PB-Cre4, Pten flox/flox ). Immunohistochemistry was performed with anti-PTEN, P-AKT (Ser473), PTK6 and P-PTK6 (Tyr342) antibodies, and samples were counterstained with DAPI (blue). The size bar denotes 20 μm. C) Prostate tumor cells with highly activated PTK6 at the plasma membrane undergo EMT. Immunohistochemistry was performed with anti-PY, P-PTK6 (PY342), CK5, CK8, p63, BrdU, Ki67, E-cadherin and vimentin antibodies, and samples were counterstained with DAPI (blue). The size bar denotes 50 μm and 20 μm, respectively.

    Journal: Cancer research

    Article Title: PTK6 activation at the membrane regulates epithelial-mesenchymal transition in prostate cancer

    doi: 10.1158/0008-5472.CAN-13-0443

    Figure Lengend Snippet: Aberrant activation of PTK6 is accompanied by deregulated E-cadherin at the plasma membrane in prostate tumor cells of PB-Cre4, Pten flox/flox mice A) An enlarged anterior prostate was observed in PB-Cre4, Pten flox/flox mice at the age of 6 months. B) Endogenous PTK6 is activated at the membrane in prostate tumor cells in a murine model (PB-Cre4, Pten flox/flox ). Immunohistochemistry was performed with anti-PTEN, P-AKT (Ser473), PTK6 and P-PTK6 (Tyr342) antibodies, and samples were counterstained with DAPI (blue). The size bar denotes 20 μm. C) Prostate tumor cells with highly activated PTK6 at the plasma membrane undergo EMT. Immunohistochemistry was performed with anti-PY, P-PTK6 (PY342), CK5, CK8, p63, BrdU, Ki67, E-cadherin and vimentin antibodies, and samples were counterstained with DAPI (blue). The size bar denotes 50 μm and 20 μm, respectively.

    Article Snippet: Anti-β-actin (AC-15) and vimentin antibodies were purchased from Sigma-Aldrich.

    Techniques: Activation Assay, Mouse Assay, Immunohistochemistry