anti-vimentin Search Results


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  • 99
    Millipore anti vimentin
    Characterization of MMCs. The isolated mesangial cells showed the expected elongated stellate shape ( A and B ). An antibody against collagen VIII–stained wild-type ( C ) but not Col8a1 − /Col8a2 − MMCs ( D ), whereas EGFP was stained in Col8a1 − /Col8a2 − ( F ) but not wild-type MMCs ( E ). Intermediate filaments typical for MMCs stained positive: <t>vimentin</t> ( G and H ) and desmin ( K and L ). An antibody against cytoskeletal α–smooth muscle actin distinguished MMCs from fibroblasts ( I and J ). Staining MMCs with antibodies against markers specific for vascular endothelial cells such as CD31/PECAM-1 was negative ( M and N ). An isotype matched mouse IgG was used as control ( O ). Wild-type: A, C, E, G, I, K, M , and O ; Col8a1 − /Col8a2 − : B, D, F, H, J, L , and N . Original magnification: ×200. (A high-quality digital representation of this figure is available in the online issue.)
    Anti Vimentin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1412 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Millipore goat anti vimentin
    U87 glioma xenograft slice culture characterization and drug responses. a Retention of tissue integrity in U87 flank xenograft slice cultures by H E staining, cell proliferation detected by Ki-67 immunostaining (red, DAPI counterstain) and cell death by SYTOX Green (SG)/Hoechst (blue) staining or by cleaved caspase 3 (CC3)/hematoxylin staining. b Xenograft slices contain endothelial (CD31), immune (CD45) and macrophage (IBA-1) cells. Confocal images identify U87 glioma cells (arrow, GFP + /HuNu + ), <t>vimentin-positive</t> mesenchymal stromal cells (vim, red, solid arrowhead) and additional non-tumor cells (open arrowhead). c – f Slices treated for 2 or 3d and assessed for cell death by SG/Hoechst staining. Tanespimycin (AAG), geldanamycin, bortezomide (BORT, 3 d), mocetinostat (MOC, 3 d), parthenolide, and YM155 (YM). Cell death was quantified by mean fluorescence (dotted regions) c , d or by % SG + /total nuclei e , f . Red outline in c is a positive control crush lesion. g , h Apoptotic cell death quantified as CC3-stained area. No primary antibody control is shown. i Cell death in U87 cells by loss of CellTox Green fluorescence 28 when grown in conventional (blue) or slice (purple) medium, versus U87 xenograft slices (black line) in slice medium (SG index). For d , f , h , average ± SEM; n = 17, 3, 6, 5, 5, 3, 3, 4, 6, 6, 3, 3, 7, 6, 9, 4, 6, 8, 6, 9, 9 slices for each drug condition, in order. One-way ANOVA versus DMSO with Dunnett’s multiple comparison test. * p
    Goat Anti Vimentin, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 133 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Atlas Antibodies anti vimentin
    Epithelial properties and mobility capability in sh-PIMT A549 cells ( A, B ) Immunoblot and intensity levels of PIMT, <t>vimentin,</t> E-cadherin, and TGFβ in sh-PIMT and sh-control A549 cells. ( C, D ) Morphologic differences between sh-PIMT and sh-control A549 cells. Scale bar, 60 μm. ( E–G ) Differences in migration and invasion capabilities between sh-PIMT and sh-control A549 cells. ( H ) Differences in proliferation rate between A549 sh-PIMT and sh-control cells. A549 cells were treated with 8.0 µg/mL of cisplatin. * indicates p
    Anti Vimentin, supplied by Atlas Antibodies, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti vimentin
    Effect of HAX-1 expression on epithelial-mesenchymal transition. (A) Western blot analysis demonstrated that HAX-1 knockdown increased E-cadherin expression, and suppressed <t>vimentin</t> and N-cadherin expression in FaDu cells. (B) Quantification of the western blotting of siRNA-transfected cells. E-cadherin protein expression was increased in the siRNA group compared with the scramble group. N-cadherin protein expression was decreased in the siRNA group compared with the scramble group. Vimentin protein expression was decreased in the siRNA group compared with the scramble group. (C) Western blot analysis demonstrated that HAX-1 overexpression decreased E-cadherin expression, and promoted vimentin and N-cadherin expression in FaDu cells transfected with HAX-1 overexpression plasmid compared with the empty vector group. (D) Quantification of the western blotting of plasmid-transfected cells. E-cadherin protein expression was decreased in the HAX-1 overexpression group compared with the vector control group. N-cadherin protein expression was increased in the HAX-1 overexpression group compared with the vector control group. Vimentin protein expression was increased in the HAX-1 overexpression group compared with the vector control group. Data are presented as the mean ± standard error of the mean. *P
    Anti Vimentin, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 2496 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Characterization of MMCs. The isolated mesangial cells showed the expected elongated stellate shape ( A and B ). An antibody against collagen VIII–stained wild-type ( C ) but not Col8a1 − /Col8a2 − MMCs ( D ), whereas EGFP was stained in Col8a1 − /Col8a2 − ( F ) but not wild-type MMCs ( E ). Intermediate filaments typical for MMCs stained positive: vimentin ( G and H ) and desmin ( K and L ). An antibody against cytoskeletal α–smooth muscle actin distinguished MMCs from fibroblasts ( I and J ). Staining MMCs with antibodies against markers specific for vascular endothelial cells such as CD31/PECAM-1 was negative ( M and N ). An isotype matched mouse IgG was used as control ( O ). Wild-type: A, C, E, G, I, K, M , and O ; Col8a1 − /Col8a2 − : B, D, F, H, J, L , and N . Original magnification: ×200. (A high-quality digital representation of this figure is available in the online issue.)

    Journal: Diabetes

    Article Title: Lack of Type VIII Collagen in Mice Ameliorates Diabetic Nephropathy

    doi: 10.2337/db08-0183

    Figure Lengend Snippet: Characterization of MMCs. The isolated mesangial cells showed the expected elongated stellate shape ( A and B ). An antibody against collagen VIII–stained wild-type ( C ) but not Col8a1 − /Col8a2 − MMCs ( D ), whereas EGFP was stained in Col8a1 − /Col8a2 − ( F ) but not wild-type MMCs ( E ). Intermediate filaments typical for MMCs stained positive: vimentin ( G and H ) and desmin ( K and L ). An antibody against cytoskeletal α–smooth muscle actin distinguished MMCs from fibroblasts ( I and J ). Staining MMCs with antibodies against markers specific for vascular endothelial cells such as CD31/PECAM-1 was negative ( M and N ). An isotype matched mouse IgG was used as control ( O ). Wild-type: A, C, E, G, I, K, M , and O ; Col8a1 − /Col8a2 − : B, D, F, H, J, L , and N . Original magnification: ×200. (A high-quality digital representation of this figure is available in the online issue.)

    Article Snippet: Primary antibodies were anti–collagen VIII (Seikagaku, Tokyo, Japan), anti-EGFP (Molecular Probes), anti-vimentin, anti-desmin, anti–smooth muscle actin (Sigma), anti-CD31/PECAM-1, and anti–collagen IV (ICN, Northeim, Germany).

    Techniques: Isolation, Staining

    Expression of protein markers of enriched cultures at passage 3 after a 14-day culture period. Immunocytochemistry demonstrated the co-expression of nestin and fibronectin and vimentin in isolated SKPs. The majority of the isolated SKPs expressed on average

    Journal: Stem Cell Investigation

    Article Title: Enrichment of skin-derived neural precursor cells from dermal cell populations by altering culture conditions

    doi: 10.21037/sci.2016.10.10

    Figure Lengend Snippet: Expression of protein markers of enriched cultures at passage 3 after a 14-day culture period. Immunocytochemistry demonstrated the co-expression of nestin and fibronectin and vimentin in isolated SKPs. The majority of the isolated SKPs expressed on average

    Article Snippet: Thereafter, cells were blocked by 3% Bovine serum albumin for 2 h (Sigma, NY, USA) and incubated with the following primary antibodies for 2 h at 4 °C: monoclonal anti-nestin, monoclonal anti-fibronectin, monoclonal anti-vimentin, monoclonal anti-βIII tubulin, monoclonal anti-GFAP, and monoclonal anti-myosin (fast skeletal, 1:100) (Sigma, NY, USA), Then, cells were rinsed with PBS three times and incubated with goat anti-mouse FITC conjugated secondary antibody (1:150) (Sigma, NY, USA) for 2 h at room temperature in darkness.

    Techniques: Expressing, Immunocytochemistry, Isolation

    Expression of protein markers of control cultures at passage 3 after a 14-day culture period. Isolated cells were absolutely positive for vimentin, weakly positive for fibronectin whereas they did not express nestin as compared to enriched culture. Scale

    Journal: Stem Cell Investigation

    Article Title: Enrichment of skin-derived neural precursor cells from dermal cell populations by altering culture conditions

    doi: 10.21037/sci.2016.10.10

    Figure Lengend Snippet: Expression of protein markers of control cultures at passage 3 after a 14-day culture period. Isolated cells were absolutely positive for vimentin, weakly positive for fibronectin whereas they did not express nestin as compared to enriched culture. Scale

    Article Snippet: Thereafter, cells were blocked by 3% Bovine serum albumin for 2 h (Sigma, NY, USA) and incubated with the following primary antibodies for 2 h at 4 °C: monoclonal anti-nestin, monoclonal anti-fibronectin, monoclonal anti-vimentin, monoclonal anti-βIII tubulin, monoclonal anti-GFAP, and monoclonal anti-myosin (fast skeletal, 1:100) (Sigma, NY, USA), Then, cells were rinsed with PBS three times and incubated with goat anti-mouse FITC conjugated secondary antibody (1:150) (Sigma, NY, USA) for 2 h at room temperature in darkness.

    Techniques: Expressing, Isolation

    Characterization of lentivirus transduction-induced ChR2 expression in cardiomyocytes and fibroblasts. ( A ) NRVM culture stained with anti-sarcomeric alpha-actinin antibody; and ( B ) NRVM culture stained with anti-vimentin antibody. n = 4 per group.

    Journal: Scientific Reports

    Article Title: Electrophysiological Properties and Viability of Neonatal Rat Ventricular Myocyte Cultures with Inducible ChR2 Expression

    doi: 10.1038/s41598-017-01723-2

    Figure Lengend Snippet: Characterization of lentivirus transduction-induced ChR2 expression in cardiomyocytes and fibroblasts. ( A ) NRVM culture stained with anti-sarcomeric alpha-actinin antibody; and ( B ) NRVM culture stained with anti-vimentin antibody. n = 4 per group.

    Article Snippet: The anti-vimentin antibody, the anti-sarcomeric-α-actinin antibody, and all other reagents were from Sigma-Aldrich (St. Louis, MO).

    Techniques: Transduction, Expressing, Staining

    Representative immunohistochemical staining of TPF EBs for cytokeratin (A), human chorionic gonadotrophin (C), and vimentin (E) and of control EBs for cytokeratin (B), human chorionic gonadotrophin (D), and vimentin (F). Panels G and H depict nonspecific

    Journal: Reproductive sciences (Thousand Oaks, Calif.)

    Article Title: Placental-Derived Mesenchyme Influences Chorionic Gonadotropin and Progesterone Secretion of Human Embryonic Stem Cell-Derived Trophoblasts

    doi: 10.1177/1933719110371853

    Figure Lengend Snippet: Representative immunohistochemical staining of TPF EBs for cytokeratin (A), human chorionic gonadotrophin (C), and vimentin (E) and of control EBs for cytokeratin (B), human chorionic gonadotrophin (D), and vimentin (F). Panels G and H depict nonspecific

    Article Snippet: Cells at 0.3 × 106 per sample were incubated with the anti-vimentin mAb (Sigma) or an isotype control for 30 minutes at 4°C.

    Techniques: Immunohistochemistry, Staining

    Representative immunohistochemical staining CI2F EBs for cytokeratin (A), human chorionic gonadotrophin (C), and vimentin (E) and of control EBs for cytokeratin (B), human chorionic gonadotrophin (D), and vimentin (F). Panels G and H depict nonspecific

    Journal: Reproductive sciences (Thousand Oaks, Calif.)

    Article Title: Placental-Derived Mesenchyme Influences Chorionic Gonadotropin and Progesterone Secretion of Human Embryonic Stem Cell-Derived Trophoblasts

    doi: 10.1177/1933719110371853

    Figure Lengend Snippet: Representative immunohistochemical staining CI2F EBs for cytokeratin (A), human chorionic gonadotrophin (C), and vimentin (E) and of control EBs for cytokeratin (B), human chorionic gonadotrophin (D), and vimentin (F). Panels G and H depict nonspecific

    Article Snippet: Cells at 0.3 × 106 per sample were incubated with the anti-vimentin mAb (Sigma) or an isotype control for 30 minutes at 4°C.

    Techniques: Immunohistochemistry, Staining

    Immunohistochemical staining of term placental fibroblasts (TPFs) for fibroblast surface protein (A), vimentin (B), and cytokeratin (C). Panel D depicts a nonspecific isotype-matched IgG negative control. TPF (top panels) and choriocarcinoma cell line,

    Journal: Reproductive sciences (Thousand Oaks, Calif.)

    Article Title: Placental-Derived Mesenchyme Influences Chorionic Gonadotropin and Progesterone Secretion of Human Embryonic Stem Cell-Derived Trophoblasts

    doi: 10.1177/1933719110371853

    Figure Lengend Snippet: Immunohistochemical staining of term placental fibroblasts (TPFs) for fibroblast surface protein (A), vimentin (B), and cytokeratin (C). Panel D depicts a nonspecific isotype-matched IgG negative control. TPF (top panels) and choriocarcinoma cell line,

    Article Snippet: Cells at 0.3 × 106 per sample were incubated with the anti-vimentin mAb (Sigma) or an isotype control for 30 minutes at 4°C.

    Techniques: Immunohistochemistry, Staining, Negative Control

    Immunocharacterization of mixed primary cultures enriched in hypothalamic neurons. Neurons obtained from rat hypothalamus at 19 days of gestation were cultured for 2 weeks. (a–i) Representative confocal images depicting MAP2 (a, green), Vimentin (b, e, and h, red), Neurofilaments (d, green), and N-CAM (g, green). Nuclei were stained with TOPRO-3 (blue). Triple labeling shows that the cultures are enriched in neurons (c, f and i). Scale bar: 80 µm.

    Journal: PLoS ONE

    Article Title: MCT2 Expression and Lactate Influx in Anorexigenic and Orexigenic Neurons of the Arcuate Nucleus

    doi: 10.1371/journal.pone.0062532

    Figure Lengend Snippet: Immunocharacterization of mixed primary cultures enriched in hypothalamic neurons. Neurons obtained from rat hypothalamus at 19 days of gestation were cultured for 2 weeks. (a–i) Representative confocal images depicting MAP2 (a, green), Vimentin (b, e, and h, red), Neurofilaments (d, green), and N-CAM (g, green). Nuclei were stained with TOPRO-3 (blue). Triple labeling shows that the cultures are enriched in neurons (c, f and i). Scale bar: 80 µm.

    Article Snippet: The immunocytochemistry was performed as previously described , using the following primary antibodies: mouse anti-MAP2 (1∶200, Millipore, Temecula, CA, USA), chicken anti-Vimentin (1∶200, Millipore), mouse anti-neurofilaments (1∶1, Hibrydoma Data Bank), mouse anti-N-CAM (1∶1, Hibrydoma Data Bank), rabbit anti-POMC (1∶200, Phoenix Pharmaceuticals, Burlingame, CA, USA), goat anti-CART (1∶200, R & D Systems, MN, USA), goat anti-AgRP (1∶200, R & D Systems) and rabbit anti-NPY (Millipore), chicken anti-MCT1 (1∶100, Millipore), chicken anti-MCT2 (1∶20, Millipore), and rabbit anti-MCT4 (1∶100, Millipore).

    Techniques: Cell Culture, Chick Chorioallantoic Membrane Assay, Staining, Labeling

    MCT2 is localized in the arcuate nucleus. (a) Schematic representation of the hypothalamic area shown in b–d. (b) Rat frontal brain section using anti-vimentin (red) and anti-GLUT1 (blue) antibodies, markers of glial cells. (c) MCT2 localization. (d) Rat frontal brain section using anti-vimentin (red), anti-GLUT1 (blue), and anti-MCT2 (green) antibodies. (e–h) MCT2 was observed in cellular membranes (e) of dorsal peri-ventricular arcuate nucleus areas; this area was negative for vimentin (f, h) and GLUT1 (g, h). GLUT1 and vimentin are co-distributed in the apical region of α-tanycytes (h, triple-arrows). (i–l) In the ventral peri-ventricular area, which contains β1-tanycytes, MCT2 was expressed in cellular membranes; this area was negative for vimentin (j) and GLUT1 (k). GLUT1 and vimentin are co-distributed in apical region of β1-tanycytes (arrows). (m–p) MCT2 was observed in the cellular membranes of β1-tanycyte processes in the ventral lateral area of the arcuate nucleus, which was negative for vimentin (n, p) and GLUT1(o, p). AN: arcuate nucleus, III–V: third ventricle, ME: median eminence. Scale bar: (b and e) 150 µm; (c–d and f–u) 50 µm.

    Journal: PLoS ONE

    Article Title: MCT2 Expression and Lactate Influx in Anorexigenic and Orexigenic Neurons of the Arcuate Nucleus

    doi: 10.1371/journal.pone.0062532

    Figure Lengend Snippet: MCT2 is localized in the arcuate nucleus. (a) Schematic representation of the hypothalamic area shown in b–d. (b) Rat frontal brain section using anti-vimentin (red) and anti-GLUT1 (blue) antibodies, markers of glial cells. (c) MCT2 localization. (d) Rat frontal brain section using anti-vimentin (red), anti-GLUT1 (blue), and anti-MCT2 (green) antibodies. (e–h) MCT2 was observed in cellular membranes (e) of dorsal peri-ventricular arcuate nucleus areas; this area was negative for vimentin (f, h) and GLUT1 (g, h). GLUT1 and vimentin are co-distributed in the apical region of α-tanycytes (h, triple-arrows). (i–l) In the ventral peri-ventricular area, which contains β1-tanycytes, MCT2 was expressed in cellular membranes; this area was negative for vimentin (j) and GLUT1 (k). GLUT1 and vimentin are co-distributed in apical region of β1-tanycytes (arrows). (m–p) MCT2 was observed in the cellular membranes of β1-tanycyte processes in the ventral lateral area of the arcuate nucleus, which was negative for vimentin (n, p) and GLUT1(o, p). AN: arcuate nucleus, III–V: third ventricle, ME: median eminence. Scale bar: (b and e) 150 µm; (c–d and f–u) 50 µm.

    Article Snippet: The immunocytochemistry was performed as previously described , using the following primary antibodies: mouse anti-MAP2 (1∶200, Millipore, Temecula, CA, USA), chicken anti-Vimentin (1∶200, Millipore), mouse anti-neurofilaments (1∶1, Hibrydoma Data Bank), mouse anti-N-CAM (1∶1, Hibrydoma Data Bank), rabbit anti-POMC (1∶200, Phoenix Pharmaceuticals, Burlingame, CA, USA), goat anti-CART (1∶200, R & D Systems, MN, USA), goat anti-AgRP (1∶200, R & D Systems) and rabbit anti-NPY (Millipore), chicken anti-MCT1 (1∶100, Millipore), chicken anti-MCT2 (1∶20, Millipore), and rabbit anti-MCT4 (1∶100, Millipore).

    Techniques:

    MCT2 is expressed in arcuate nucleus neurons. (a–c) Low magnification analysis of the basal hypothalamic area using anti-vimentin (a, orange), anti-GFAP (b, red) anti-MCT2 (c, green) antibodies. TOPRO-3 was used as nuclear stain (blue). (d) Schematic representation of the hypothalamic area shown in e. (e) High magnification analysis using quadruple labeling. MCT2 labeling was negative in β1v-tanycytes (positive for vimentin) or β1d-tanycytes (positive for GFAP) or astrocytes (positive for GFAP). The MCT2 reaction was concentrated in membranes of arcuate neurons (e, arrows), and neuronal processes (e, head arrows). AN: arcuate nucleus, III–V: third ventricle, ME: median eminence, n: neurons. Scale bar: 10 µm.

    Journal: PLoS ONE

    Article Title: MCT2 Expression and Lactate Influx in Anorexigenic and Orexigenic Neurons of the Arcuate Nucleus

    doi: 10.1371/journal.pone.0062532

    Figure Lengend Snippet: MCT2 is expressed in arcuate nucleus neurons. (a–c) Low magnification analysis of the basal hypothalamic area using anti-vimentin (a, orange), anti-GFAP (b, red) anti-MCT2 (c, green) antibodies. TOPRO-3 was used as nuclear stain (blue). (d) Schematic representation of the hypothalamic area shown in e. (e) High magnification analysis using quadruple labeling. MCT2 labeling was negative in β1v-tanycytes (positive for vimentin) or β1d-tanycytes (positive for GFAP) or astrocytes (positive for GFAP). The MCT2 reaction was concentrated in membranes of arcuate neurons (e, arrows), and neuronal processes (e, head arrows). AN: arcuate nucleus, III–V: third ventricle, ME: median eminence, n: neurons. Scale bar: 10 µm.

    Article Snippet: The immunocytochemistry was performed as previously described , using the following primary antibodies: mouse anti-MAP2 (1∶200, Millipore, Temecula, CA, USA), chicken anti-Vimentin (1∶200, Millipore), mouse anti-neurofilaments (1∶1, Hibrydoma Data Bank), mouse anti-N-CAM (1∶1, Hibrydoma Data Bank), rabbit anti-POMC (1∶200, Phoenix Pharmaceuticals, Burlingame, CA, USA), goat anti-CART (1∶200, R & D Systems, MN, USA), goat anti-AgRP (1∶200, R & D Systems) and rabbit anti-NPY (Millipore), chicken anti-MCT1 (1∶100, Millipore), chicken anti-MCT2 (1∶20, Millipore), and rabbit anti-MCT4 (1∶100, Millipore).

    Techniques: Staining, Labeling

    HIF2α stabilization causes pancreatic abnormalities reminiscent of pancreatitis. Gomori trichome staining reveals increased connective tissue (stained in blue) in Pdx1-Cre;HIF2dPA pancreata ( B ) compared to control pancreata ( A ) at 8 weeks of age. Increase in smooth muscle actin (SMA) positive cells (white arrowhead) in Pdx1-Cre;HIF2dPA pancreata ( D ) compared to control pancreata ( C ) at 8 weeks of age. Ducts are marked by staining with lectin Dolichos biflorus agglutinin (DBA). Increase of the mesenchymal marker vimentin in Pdx1-Cre;HIF2dPA pancreata ( F , black arrowheads) at 8 weeks of age. Only a few scattered cells are positive for vimentin in control pancreata ( E , black arrowhead). ( G ) Quantification of vimentin-positive area per total pancreatic area. CD11b immunohistochemistry in control ( H ) and Pdx1-Cre;HIF2dPA pancreata ( I , black arrowheads) at eight weeks of age. ( J ) Quantification of CD11b-positive cells per field. F4/80 immunohistochemistry in control ( K ) and Pdx1-Cre;HIF2dPA pancreata ( L , black arrowheads) at eight weeks of age. ( M ) Quantification of F4/80-positive cells per field. Data points represent values for each individual mouse. The mean value is indicated as a horizontal line. Scale bars = 50 µm. ** P

    Journal: Scientific Reports

    Article Title: Stabilization of HIF-2α impacts pancreas growth

    doi: 10.1038/s41598-018-32054-5

    Figure Lengend Snippet: HIF2α stabilization causes pancreatic abnormalities reminiscent of pancreatitis. Gomori trichome staining reveals increased connective tissue (stained in blue) in Pdx1-Cre;HIF2dPA pancreata ( B ) compared to control pancreata ( A ) at 8 weeks of age. Increase in smooth muscle actin (SMA) positive cells (white arrowhead) in Pdx1-Cre;HIF2dPA pancreata ( D ) compared to control pancreata ( C ) at 8 weeks of age. Ducts are marked by staining with lectin Dolichos biflorus agglutinin (DBA). Increase of the mesenchymal marker vimentin in Pdx1-Cre;HIF2dPA pancreata ( F , black arrowheads) at 8 weeks of age. Only a few scattered cells are positive for vimentin in control pancreata ( E , black arrowhead). ( G ) Quantification of vimentin-positive area per total pancreatic area. CD11b immunohistochemistry in control ( H ) and Pdx1-Cre;HIF2dPA pancreata ( I , black arrowheads) at eight weeks of age. ( J ) Quantification of CD11b-positive cells per field. F4/80 immunohistochemistry in control ( K ) and Pdx1-Cre;HIF2dPA pancreata ( L , black arrowheads) at eight weeks of age. ( M ) Quantification of F4/80-positive cells per field. Data points represent values for each individual mouse. The mean value is indicated as a horizontal line. Scale bars = 50 µm. ** P

    Article Snippet: Primary antibodies were used at the indicated dilution: Rabbit anti-cleaved caspase-3 (1:50, 9661), rabbit anti-phosphorylated-ERK (1:300, 4370) and rabbit anti-phosphorylated-S6 ribosomal protein (1:75, 4858) from Cell Signaling Technology; rabbit anti-carboxypeptidase A (1:200, 1810-0006) from AbD Serotec; mouse anti-E-cadherin (1:200, 610181), rat anti-panendothelial cell antigen (MECA32) (1:300, 553849) and mouse anti-β-catenin (1:300, 610153) from BD Pharmigen; rat anti-F4/80 (1:50, T-2006; BMA Biomedicals); rat anti-cytokeratin 19 (1:100, TROMA-III; Developmental Studies Hybridoma Bank); rabbit anti-ZO-1 (1:100, 40-2300; Invitrogen); rabbit anti-Sox-9 (1:3000, AB5535; Merck Millipore); rabbit anti-CD11b (1:200, NB110-89474) and rabbit anti-HIF2α diluted 1:100 (NB100-122) from Novus; mouse anti-amylase (1:200; Sc-46657) goat anti-α-clusterin (1:200, sc-6419) and rabbit anti-PKCε (1:100, sc-214) from Santa Cruz Biotechnology; mouse anti-acetylated tubulin (1:10000, T-6793), mouse anti-α-smooth muscle actin (1:1000, A-5228), rabbit anti-laminin (1:500, L-9393) and rabbit anti-vimentin (1:1000, AB92547) from Sigma-Aldrich and armenian hamster anti-Mucin-1 (1:200, HM-1630; Thermo Scientific).

    Techniques: Staining, Marker, Immunohistochemistry

    Subplate neuron cultures contain highly purified, glutamatergic neurons but few synapses. ( A ) Subplate cultures immunostained at 7 d.i.v. for Tau (pseudocolored green) to label neurons and vimentin (red) to label nonneuronal cells. ( B ) Subplate cultures immunostained for glutamate (green) with a Hoescht counterstain for nuclei (blue). ( C ) Immunofluorescence for GAD-65 and GAD-67 (combined in red) to label GABAergic neurons, with all neurons counterstained for Tau (green). ( D ) Subplate or ( E ) hippocampal neurons cultured for 10 days then immunostained for synapsin (red) and PSD-95 (green); counterstained using phalloidin to detect actin (blue). Synapses are defined as puncta colocalizing synapsin and PSD-95 (arrowheads), which appear yellow (or white in the presence of the blue phalloidin counterstain). Inset: magnified view of a portion of dendrite from each image. ( F ) Quantification of colocalized puncta per square micron of phalloidin-labeled neurite in subplate versus hippocampal neurons at indicated times in culture. Mean ± SEM, compiled from 3 experiments. Asterisks = P

    Journal: Cerebral Cortex (New York, NY)

    Article Title: Synaptogenesis in Purified Cortical Subplate Neurons

    doi: 10.1093/cercor/bhn194

    Figure Lengend Snippet: Subplate neuron cultures contain highly purified, glutamatergic neurons but few synapses. ( A ) Subplate cultures immunostained at 7 d.i.v. for Tau (pseudocolored green) to label neurons and vimentin (red) to label nonneuronal cells. ( B ) Subplate cultures immunostained for glutamate (green) with a Hoescht counterstain for nuclei (blue). ( C ) Immunofluorescence for GAD-65 and GAD-67 (combined in red) to label GABAergic neurons, with all neurons counterstained for Tau (green). ( D ) Subplate or ( E ) hippocampal neurons cultured for 10 days then immunostained for synapsin (red) and PSD-95 (green); counterstained using phalloidin to detect actin (blue). Synapses are defined as puncta colocalizing synapsin and PSD-95 (arrowheads), which appear yellow (or white in the presence of the blue phalloidin counterstain). Inset: magnified view of a portion of dendrite from each image. ( F ) Quantification of colocalized puncta per square micron of phalloidin-labeled neurite in subplate versus hippocampal neurons at indicated times in culture. Mean ± SEM, compiled from 3 experiments. Asterisks = P

    Article Snippet: Antibodies used in other experiments were rabbit anti-Tau (Sigma), mouse anti-vimentin (Chemicon), rabbit anti-glutamate (Chemicon), mouse anti-GAD65 (Chemicon), mouse anti-GAD67 (a gift of Dr Daniel Kaufman, UCLA), rabbit anti-GluR1 (Chemicon), and goat anti-GFP (Abcam, Cambridge, UK).

    Techniques: Purification, Immunofluorescence, Cell Culture, Labeling

    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

    Retinal wholemounts stained for GFAP (red) and vimentin (green) demonstrate both Müller cells and astrocytes in preretinal glial structures

    Journal: Experimental eye research

    Article Title: Idiopathic preretinal glia in aging and age-related macular degeneration

    doi: 10.1016/j.exer.2015.07.016

    Figure Lengend Snippet: Retinal wholemounts stained for GFAP (red) and vimentin (green) demonstrate both Müller cells and astrocytes in preretinal glial structures

    Article Snippet: Primary antibodies used on cross sections included: rabbit anti-pan laminin (Sigma, 1:200) and chick anti-vimentin (Millipore, 1:500).

    Techniques: Staining

    Micrographs of oviductal epithelial (A) and stromal cells (B) in monolayer culture and immunostaining by anti-cytokeratin and anti-vimentin antibodies in ampullary oviductal epithelial cells (C, E) and stromal cells (D, F). Anti-mouse-IgG conjugate-donkey (green) was used for staining cytokeratin and vimentin as the secondary antibody. DAPI (blue) was used to visualize nuclei. Staining in the isthmus was virtually the same as that in the ampulla. Each scale bar indicates 100 μm.

    Journal: The Journal of Reproduction and Development

    Article Title: Purified Culture Systems for Bovine Oviductal Stromal Cells

    doi: 10.1262/jrd.2013-061

    Figure Lengend Snippet: Micrographs of oviductal epithelial (A) and stromal cells (B) in monolayer culture and immunostaining by anti-cytokeratin and anti-vimentin antibodies in ampullary oviductal epithelial cells (C, E) and stromal cells (D, F). Anti-mouse-IgG conjugate-donkey (green) was used for staining cytokeratin and vimentin as the secondary antibody. DAPI (blue) was used to visualize nuclei. Staining in the isthmus was virtually the same as that in the ampulla. Each scale bar indicates 100 μm.

    Article Snippet: Evaluation of cell homogeneity The homogeneity of the epithelial cells and stromal cells was evaluated by immunofluorescent staining using anti-cytokeratin IgG produced in the mouse (Sigma-Aldrich) and anti-vimentin IgG produced in the mouse (Sigma-Aldrich) as the primary antibodies, anti-mouse IgG (FITC) conjugate-donkey (Sigma-Aldrich) as the secondary antibody, and ProLong Gold Antifade Reagent with DAPI (Invitrogen) as described previously with our modification [ , ].

    Techniques: Immunostaining, Staining

    Nupr1 silencing triggers OIS in human pancreatic cancer cells. ( a ) Western blot and RT-qPCR of Nupr1 showing the effectiveness of specific siRNA in MiaPaCa2 cells. ( b ) Cytometric cell cycle analysis showing increase of G1-arrested cells by siNupr1 treatment in MiaPaCa2 cells. ( c ) MiaPaCa2 cell proliferation curve shows a significant stop in cell proliferation after siNupr1 silencing compared with siControl-treated cells. The relative cell number at each time point on the growth curves represents the mean±S.D. of triplicate normalized to the cell number at day 1. ( d ) SA- β Gal staining of MiaPaca2 cells. A very significant increase of senescent cells is observed after Nupr1 depletion. ( e ) SA- β Gal activity staining in Panc1, CaPan-2 and BxPC-3 cells treated with a siRNA against Nupr1. ( f ) Bright field and β -tubulin immunofluorescence of MiaPaCa2 cells treated with siNupr1 or siControl. Change in cell morphology and significant increase in cell size is observed in Nupr1-silenced cells. ( g ) Vimentin immunofluorescence showing significant increase of focal adhesions number in siNupr1-treated cells. Error bars±S.D.; * P

    Journal: Cell Death and Differentiation

    Article Title: Genetic inactivation of the pancreatitis-inducible gene Nupr1 impairs PanIN formation by modulating KrasG12D-induced senescence

    doi: 10.1038/cdd.2014.74

    Figure Lengend Snippet: Nupr1 silencing triggers OIS in human pancreatic cancer cells. ( a ) Western blot and RT-qPCR of Nupr1 showing the effectiveness of specific siRNA in MiaPaCa2 cells. ( b ) Cytometric cell cycle analysis showing increase of G1-arrested cells by siNupr1 treatment in MiaPaCa2 cells. ( c ) MiaPaCa2 cell proliferation curve shows a significant stop in cell proliferation after siNupr1 silencing compared with siControl-treated cells. The relative cell number at each time point on the growth curves represents the mean±S.D. of triplicate normalized to the cell number at day 1. ( d ) SA- β Gal staining of MiaPaca2 cells. A very significant increase of senescent cells is observed after Nupr1 depletion. ( e ) SA- β Gal activity staining in Panc1, CaPan-2 and BxPC-3 cells treated with a siRNA against Nupr1. ( f ) Bright field and β -tubulin immunofluorescence of MiaPaCa2 cells treated with siNupr1 or siControl. Change in cell morphology and significant increase in cell size is observed in Nupr1-silenced cells. ( g ) Vimentin immunofluorescence showing significant increase of focal adhesions number in siNupr1-treated cells. Error bars±S.D.; * P

    Article Snippet: Primary antibodies used were as follows: anti-p27, anti-Cdk2, anti-Cdk4, anti-cyclin D1 and anti-Rb polyclonal antibodies were from Santa Cruz Biotechnologies; anti-Nupr1 monoclonal antibody was homemade and described previously; FoxO3a monoclonal, phospho-FoxO3a (Ser253) polyclonal and phospho-Rb (Ser807/811) monoclonal antibodies were from Cell Signaling; anti-β -tubulin polyclonal and anti-vimentin monoclonal antibodies were from Sigma-Aldrich.

    Techniques: Western Blot, Quantitative RT-PCR, Cell Cycle Assay, Staining, Activity Assay, Immunofluorescence

    U87 glioma xenograft slice culture characterization and drug responses. a Retention of tissue integrity in U87 flank xenograft slice cultures by H E staining, cell proliferation detected by Ki-67 immunostaining (red, DAPI counterstain) and cell death by SYTOX Green (SG)/Hoechst (blue) staining or by cleaved caspase 3 (CC3)/hematoxylin staining. b Xenograft slices contain endothelial (CD31), immune (CD45) and macrophage (IBA-1) cells. Confocal images identify U87 glioma cells (arrow, GFP + /HuNu + ), vimentin-positive mesenchymal stromal cells (vim, red, solid arrowhead) and additional non-tumor cells (open arrowhead). c – f Slices treated for 2 or 3d and assessed for cell death by SG/Hoechst staining. Tanespimycin (AAG), geldanamycin, bortezomide (BORT, 3 d), mocetinostat (MOC, 3 d), parthenolide, and YM155 (YM). Cell death was quantified by mean fluorescence (dotted regions) c , d or by % SG + /total nuclei e , f . Red outline in c is a positive control crush lesion. g , h Apoptotic cell death quantified as CC3-stained area. No primary antibody control is shown. i Cell death in U87 cells by loss of CellTox Green fluorescence 28 when grown in conventional (blue) or slice (purple) medium, versus U87 xenograft slices (black line) in slice medium (SG index). For d , f , h , average ± SEM; n = 17, 3, 6, 5, 5, 3, 3, 4, 6, 6, 3, 3, 7, 6, 9, 4, 6, 8, 6, 9, 9 slices for each drug condition, in order. One-way ANOVA versus DMSO with Dunnett’s multiple comparison test. * p

    Journal: NPJ Precision Oncology

    Article Title: Multiplexed drug testing of tumor slices using a microfluidic platform

    doi: 10.1038/s41698-020-0117-y

    Figure Lengend Snippet: U87 glioma xenograft slice culture characterization and drug responses. a Retention of tissue integrity in U87 flank xenograft slice cultures by H E staining, cell proliferation detected by Ki-67 immunostaining (red, DAPI counterstain) and cell death by SYTOX Green (SG)/Hoechst (blue) staining or by cleaved caspase 3 (CC3)/hematoxylin staining. b Xenograft slices contain endothelial (CD31), immune (CD45) and macrophage (IBA-1) cells. Confocal images identify U87 glioma cells (arrow, GFP + /HuNu + ), vimentin-positive mesenchymal stromal cells (vim, red, solid arrowhead) and additional non-tumor cells (open arrowhead). c – f Slices treated for 2 or 3d and assessed for cell death by SG/Hoechst staining. Tanespimycin (AAG), geldanamycin, bortezomide (BORT, 3 d), mocetinostat (MOC, 3 d), parthenolide, and YM155 (YM). Cell death was quantified by mean fluorescence (dotted regions) c , d or by % SG + /total nuclei e , f . Red outline in c is a positive control crush lesion. g , h Apoptotic cell death quantified as CC3-stained area. No primary antibody control is shown. i Cell death in U87 cells by loss of CellTox Green fluorescence 28 when grown in conventional (blue) or slice (purple) medium, versus U87 xenograft slices (black line) in slice medium (SG index). For d , f , h , average ± SEM; n = 17, 3, 6, 5, 5, 3, 3, 4, 6, 6, 3, 3, 7, 6, 9, 4, 6, 8, 6, 9, 9 slices for each drug condition, in order. One-way ANOVA versus DMSO with Dunnett’s multiple comparison test. * p

    Article Snippet: Tissue was incubated with primary antibody (diluted in TNB) overnight at 4 C. Primary antibodies were as follows: rabbit anti-Ki-67 (1/300 fluorescence 1/4000 DAB), AbCAM, ab15580), rabbit anti-CD31 (1/60, AbCAM ab28364), rabbit anti-Iba-1 (1/1000, WAKO), rabbit anti-active caspase 3 (Asp175) (1/600, Cell Signaling, Antibody #9661), mouse anti-HuNu (1/500, AbCAM ab191181), goat anti-vimentin (1/300, Chemicon AB1620), rabbit anti-CD45 (1/1000, AbCAM, ab10558).

    Techniques: Staining, Immunostaining, Fluorescence, Positive Control

    Epithelial properties and mobility capability in sh-PIMT A549 cells ( A, B ) Immunoblot and intensity levels of PIMT, vimentin, E-cadherin, and TGFβ in sh-PIMT and sh-control A549 cells. ( C, D ) Morphologic differences between sh-PIMT and sh-control A549 cells. Scale bar, 60 μm. ( E–G ) Differences in migration and invasion capabilities between sh-PIMT and sh-control A549 cells. ( H ) Differences in proliferation rate between A549 sh-PIMT and sh-control cells. A549 cells were treated with 8.0 µg/mL of cisplatin. * indicates p

    Journal: Oncotarget

    Article Title: Deficiency of protein-L-isoaspartate (D-aspartate) O-methyl-transferase expression under endoplasmic reticulum stress promotes epithelial mesenchymal transition in lung adenocarcinoma

    doi: 10.18632/oncotarget.24324

    Figure Lengend Snippet: Epithelial properties and mobility capability in sh-PIMT A549 cells ( A, B ) Immunoblot and intensity levels of PIMT, vimentin, E-cadherin, and TGFβ in sh-PIMT and sh-control A549 cells. ( C, D ) Morphologic differences between sh-PIMT and sh-control A549 cells. Scale bar, 60 μm. ( E–G ) Differences in migration and invasion capabilities between sh-PIMT and sh-control A549 cells. ( H ) Differences in proliferation rate between A549 sh-PIMT and sh-control cells. A549 cells were treated with 8.0 µg/mL of cisplatin. * indicates p

    Article Snippet: Antibodies and reagents Anti-PIMT (Abcam PLC, Cambridge, UK, 97446, 1:2000 dilution), anti-E-cadherin (Santa Cruz Biotechnology Inc., Santa Cruz, USA, sc-7870, 1:200 dilution), anti-vimentin (sc-7557, 1:250 dilution), anti-α tubulin (sc-5546, 1:400 dilution), anti-GRP78 (sc-1051, 1:200 dilution), anti-p53 (sc-1616, 1:200 dilution), anti-β actin (sc-32293, 1:200 dilution), anti-GAPDH (sc-25778), anti-Zeb1 (Atlas Antibodies AB, Stockholm, Sweden, HPA027524, 1:500 dilution), anti-Slug (sc-15391, 1:200 dilution), anti-Twist (Proteintech, Rosemont, IL, USA, 1:500 dilution), anti-Snail1 (Proteintech, 1:1000 dilution), anti-HIF1α (Novus Biologicals, Minneapolis, USA, NB100-479, 1:1000 dilution), and anti-HIF1α (Abcam, ab51608, 1:1000 dilution) antibodies were used in this study.

    Techniques: Migration

    Response of lung adenocarcinoma cell lines to Thapsigargin ( A ) Immunoblotting of GRP78, PIMT, p53, vimentin, and E-cadherin in A549 cells treated with Thapsigargin (Tg). Line 1, DMSO; Line 2, 1.0 × 10 –4 µM of Tg; Line 3, 1.0 × 10 –3 µM of Tg; Line 4, 5.0 × 10 –3 µM of Tg; Line 5, 1.0 × 10 –2 µM of Tg; Line 6, 5.0 × 10 –2 µM of Tg; Line 7, 0.1 µM of Tg; Line 8, 0.2 µM of Tg; Line 9, 0.5 µM of Tg. ( B ) Intensity of GRP78 and p53 in A549 cells treated with Tg. ( C ) Intensity of PIMT and E-cadherin in A549 cells treated with Tg. Immunoblotting and relative intensity of GRP78, PIMT, vimentin, and E-cadherin in H441 ( D–F ), H1650 ( G–I ), and H1650 ( J–L ) cells treated with Tg. Line 1, DMSO; Line 2, 1.0 × 10 –4 µM of Tg; Line 3, 1.0 × 10 –3 µM of Tg; Line 4, 1.0 × 10 –2 µM of Tg; Line 5, 0.1 µM of Tg; Line 6, 1.0 µM of Tg.

    Journal: Oncotarget

    Article Title: Deficiency of protein-L-isoaspartate (D-aspartate) O-methyl-transferase expression under endoplasmic reticulum stress promotes epithelial mesenchymal transition in lung adenocarcinoma

    doi: 10.18632/oncotarget.24324

    Figure Lengend Snippet: Response of lung adenocarcinoma cell lines to Thapsigargin ( A ) Immunoblotting of GRP78, PIMT, p53, vimentin, and E-cadherin in A549 cells treated with Thapsigargin (Tg). Line 1, DMSO; Line 2, 1.0 × 10 –4 µM of Tg; Line 3, 1.0 × 10 –3 µM of Tg; Line 4, 5.0 × 10 –3 µM of Tg; Line 5, 1.0 × 10 –2 µM of Tg; Line 6, 5.0 × 10 –2 µM of Tg; Line 7, 0.1 µM of Tg; Line 8, 0.2 µM of Tg; Line 9, 0.5 µM of Tg. ( B ) Intensity of GRP78 and p53 in A549 cells treated with Tg. ( C ) Intensity of PIMT and E-cadherin in A549 cells treated with Tg. Immunoblotting and relative intensity of GRP78, PIMT, vimentin, and E-cadherin in H441 ( D–F ), H1650 ( G–I ), and H1650 ( J–L ) cells treated with Tg. Line 1, DMSO; Line 2, 1.0 × 10 –4 µM of Tg; Line 3, 1.0 × 10 –3 µM of Tg; Line 4, 1.0 × 10 –2 µM of Tg; Line 5, 0.1 µM of Tg; Line 6, 1.0 µM of Tg.

    Article Snippet: Antibodies and reagents Anti-PIMT (Abcam PLC, Cambridge, UK, 97446, 1:2000 dilution), anti-E-cadherin (Santa Cruz Biotechnology Inc., Santa Cruz, USA, sc-7870, 1:200 dilution), anti-vimentin (sc-7557, 1:250 dilution), anti-α tubulin (sc-5546, 1:400 dilution), anti-GRP78 (sc-1051, 1:200 dilution), anti-p53 (sc-1616, 1:200 dilution), anti-β actin (sc-32293, 1:200 dilution), anti-GAPDH (sc-25778), anti-Zeb1 (Atlas Antibodies AB, Stockholm, Sweden, HPA027524, 1:500 dilution), anti-Slug (sc-15391, 1:200 dilution), anti-Twist (Proteintech, Rosemont, IL, USA, 1:500 dilution), anti-Snail1 (Proteintech, 1:1000 dilution), anti-HIF1α (Novus Biologicals, Minneapolis, USA, NB100-479, 1:1000 dilution), and anti-HIF1α (Abcam, ab51608, 1:1000 dilution) antibodies were used in this study.

    Techniques:

    Supplemental expression of PIMT reduces EMT and cancer invasion in A549 cells induced by hypoxic conditions ( A, B ) Immunoblotting and relative intensity of GRP78 in A549 cells under normal hypoxic (1% O 2 ) conditions. ( C ) Immunoblotting of PIMT, vimentin, and E-cadherin in A549 cells under normal (21% O 2 ) and hypoxic (1% O 2 ) conditions treated with and without PIMT vector. ( D–F ) Intensity of PIMT, vimentin, and E-cadherin in A549 cells treated with and without PIMT vector. Line 1: normoxia + empty vector, Line 2: hypoxia + empty vector, Line 3: normoxia + PIMT vector, Line 4: hypoxia + PIMT vector. ( G ) Migration cell count, ( H ) Invasion cell count, and ( I ) Invasion rate in A549 cells under hypoxic conditions with empty and PIMT vector. * indicates p

    Journal: Oncotarget

    Article Title: Deficiency of protein-L-isoaspartate (D-aspartate) O-methyl-transferase expression under endoplasmic reticulum stress promotes epithelial mesenchymal transition in lung adenocarcinoma

    doi: 10.18632/oncotarget.24324

    Figure Lengend Snippet: Supplemental expression of PIMT reduces EMT and cancer invasion in A549 cells induced by hypoxic conditions ( A, B ) Immunoblotting and relative intensity of GRP78 in A549 cells under normal hypoxic (1% O 2 ) conditions. ( C ) Immunoblotting of PIMT, vimentin, and E-cadherin in A549 cells under normal (21% O 2 ) and hypoxic (1% O 2 ) conditions treated with and without PIMT vector. ( D–F ) Intensity of PIMT, vimentin, and E-cadherin in A549 cells treated with and without PIMT vector. Line 1: normoxia + empty vector, Line 2: hypoxia + empty vector, Line 3: normoxia + PIMT vector, Line 4: hypoxia + PIMT vector. ( G ) Migration cell count, ( H ) Invasion cell count, and ( I ) Invasion rate in A549 cells under hypoxic conditions with empty and PIMT vector. * indicates p

    Article Snippet: Antibodies and reagents Anti-PIMT (Abcam PLC, Cambridge, UK, 97446, 1:2000 dilution), anti-E-cadherin (Santa Cruz Biotechnology Inc., Santa Cruz, USA, sc-7870, 1:200 dilution), anti-vimentin (sc-7557, 1:250 dilution), anti-α tubulin (sc-5546, 1:400 dilution), anti-GRP78 (sc-1051, 1:200 dilution), anti-p53 (sc-1616, 1:200 dilution), anti-β actin (sc-32293, 1:200 dilution), anti-GAPDH (sc-25778), anti-Zeb1 (Atlas Antibodies AB, Stockholm, Sweden, HPA027524, 1:500 dilution), anti-Slug (sc-15391, 1:200 dilution), anti-Twist (Proteintech, Rosemont, IL, USA, 1:500 dilution), anti-Snail1 (Proteintech, 1:1000 dilution), anti-HIF1α (Novus Biologicals, Minneapolis, USA, NB100-479, 1:1000 dilution), and anti-HIF1α (Abcam, ab51608, 1:1000 dilution) antibodies were used in this study.

    Techniques: Expressing, Plasmid Preparation, Migration, Cell Counting

    PIMT expression in cancer cell lines and epithelial properties in si-PIMT cancer cells ( A ) Immunoblotting of PIMT, GRP78, p53, vimentin, and E-cadherin in 6 lung adenocarcinoma cell lines: A549, H441, H460, H1650, Calu 1, and Calu 6. ( B ) Expression levels of PIMT in the six cell lines. ( C, D ) Immunoblot and intensity levels of PIMT, vimentin, and E-cadherin in A549 cells interfered by PIMT si-RNA anti-sense (J-010000-05-0002 #1 and J-010000-07-0002 #2 ). Immunoblot and intensity levels of vimentin, E-cadherin, and PIMT in H441 ( E, F) , H1650 ( G, H ), H460 ( I, J ), Calu1 ( K, L) and Calu6 cells ( M, N ) interfered by PIMT si-RNA anti-sense (J-010000-05-0002 ¶ and J-010000-07-0002 § ). * indicates p

    Journal: Oncotarget

    Article Title: Deficiency of protein-L-isoaspartate (D-aspartate) O-methyl-transferase expression under endoplasmic reticulum stress promotes epithelial mesenchymal transition in lung adenocarcinoma

    doi: 10.18632/oncotarget.24324

    Figure Lengend Snippet: PIMT expression in cancer cell lines and epithelial properties in si-PIMT cancer cells ( A ) Immunoblotting of PIMT, GRP78, p53, vimentin, and E-cadherin in 6 lung adenocarcinoma cell lines: A549, H441, H460, H1650, Calu 1, and Calu 6. ( B ) Expression levels of PIMT in the six cell lines. ( C, D ) Immunoblot and intensity levels of PIMT, vimentin, and E-cadherin in A549 cells interfered by PIMT si-RNA anti-sense (J-010000-05-0002 #1 and J-010000-07-0002 #2 ). Immunoblot and intensity levels of vimentin, E-cadherin, and PIMT in H441 ( E, F) , H1650 ( G, H ), H460 ( I, J ), Calu1 ( K, L) and Calu6 cells ( M, N ) interfered by PIMT si-RNA anti-sense (J-010000-05-0002 ¶ and J-010000-07-0002 § ). * indicates p

    Article Snippet: Antibodies and reagents Anti-PIMT (Abcam PLC, Cambridge, UK, 97446, 1:2000 dilution), anti-E-cadherin (Santa Cruz Biotechnology Inc., Santa Cruz, USA, sc-7870, 1:200 dilution), anti-vimentin (sc-7557, 1:250 dilution), anti-α tubulin (sc-5546, 1:400 dilution), anti-GRP78 (sc-1051, 1:200 dilution), anti-p53 (sc-1616, 1:200 dilution), anti-β actin (sc-32293, 1:200 dilution), anti-GAPDH (sc-25778), anti-Zeb1 (Atlas Antibodies AB, Stockholm, Sweden, HPA027524, 1:500 dilution), anti-Slug (sc-15391, 1:200 dilution), anti-Twist (Proteintech, Rosemont, IL, USA, 1:500 dilution), anti-Snail1 (Proteintech, 1:1000 dilution), anti-HIF1α (Novus Biologicals, Minneapolis, USA, NB100-479, 1:1000 dilution), and anti-HIF1α (Abcam, ab51608, 1:1000 dilution) antibodies were used in this study.

    Techniques: Expressing

    Effect of HAX-1 expression on epithelial-mesenchymal transition. (A) Western blot analysis demonstrated that HAX-1 knockdown increased E-cadherin expression, and suppressed vimentin and N-cadherin expression in FaDu cells. (B) Quantification of the western blotting of siRNA-transfected cells. E-cadherin protein expression was increased in the siRNA group compared with the scramble group. N-cadherin protein expression was decreased in the siRNA group compared with the scramble group. Vimentin protein expression was decreased in the siRNA group compared with the scramble group. (C) Western blot analysis demonstrated that HAX-1 overexpression decreased E-cadherin expression, and promoted vimentin and N-cadherin expression in FaDu cells transfected with HAX-1 overexpression plasmid compared with the empty vector group. (D) Quantification of the western blotting of plasmid-transfected cells. E-cadherin protein expression was decreased in the HAX-1 overexpression group compared with the vector control group. N-cadherin protein expression was increased in the HAX-1 overexpression group compared with the vector control group. Vimentin protein expression was increased in the HAX-1 overexpression group compared with the vector control group. Data are presented as the mean ± standard error of the mean. *P

    Journal: Molecular Medicine Reports

    Article Title: Abnormal expression of HAX-1 is associated with cellular proliferation and migration in human hypopharyngeal squamous cell carcinoma

    doi: 10.3892/mmr.2017.7155

    Figure Lengend Snippet: Effect of HAX-1 expression on epithelial-mesenchymal transition. (A) Western blot analysis demonstrated that HAX-1 knockdown increased E-cadherin expression, and suppressed vimentin and N-cadherin expression in FaDu cells. (B) Quantification of the western blotting of siRNA-transfected cells. E-cadherin protein expression was increased in the siRNA group compared with the scramble group. N-cadherin protein expression was decreased in the siRNA group compared with the scramble group. Vimentin protein expression was decreased in the siRNA group compared with the scramble group. (C) Western blot analysis demonstrated that HAX-1 overexpression decreased E-cadherin expression, and promoted vimentin and N-cadherin expression in FaDu cells transfected with HAX-1 overexpression plasmid compared with the empty vector group. (D) Quantification of the western blotting of plasmid-transfected cells. E-cadherin protein expression was decreased in the HAX-1 overexpression group compared with the vector control group. N-cadherin protein expression was increased in the HAX-1 overexpression group compared with the vector control group. Vimentin protein expression was increased in the HAX-1 overexpression group compared with the vector control group. Data are presented as the mean ± standard error of the mean. *P

    Article Snippet: Following blocking in 5% non-fat milk for 1 h at room temperature, the membrane was incubated with the following primary antibodies at 4°C overnight: Anti-HAX-1 (1:500; cat. no. sc-28268; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-E-cadherin (1:1,000; cat. no. ab1416; Abcam, Cambridge, MA, USA), anti-vimentin (1:1,000; cat. no. ab92547; Abcam), anti-N-cadherin (1:500; cat. no. ab18203; Abcam) and anti-β-actin (1:2,000; cat. no. A1978; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany).

    Techniques: Expressing, Western Blot, Transfection, Over Expression, Plasmid Preparation