human lamin b1 Search Results


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  • 93
    Abcam lamin b1
    Lamin B1, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 1942 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc human mcerulean lamin b1 10
    Human Mcerulean Lamin B1 10, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    OriGene human lamin b1 cdna
    Human Lamin B1 Cdna, supplied by OriGene, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rabbit anti human lamin b1
    Rabbit Anti Human Lamin B1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam polyclonal anti human lamin b1
    Polyclonal Anti Human Lamin B1, supplied by Abcam, 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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    80
    Matritech anti human lamin b1 mab
    Anti Human Lamin B1 Mab, supplied by Matritech, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology monoclonal anti human lamin b1
    Curcumin induces the processing of caspase-3 and the cleavage of cytoskeletal proteins. PMNs (10 × 10 6 cells/mL) were incubated for the indicated periods of time with 10 or 50 µM curcumin (Cur10 and Cur50, respectively), and (a) the processing of procaspase-3 (proCASP3) or (b) the cleavage of <t>lamin</t> B1 or vimentin (Vim) were assessed by Western blot as described in the ‘Materials and methods’ section. Results are from one representative experiment out of at least three. Arrowhead indicated the 47-kDa fragment of lamin B 1 observed in HL-60, PLB-985 and human PMNs. 22 SA: spontaneous apoptosis; PMNs: polymorphonuclear neutrophil cells; ATO: arsenic trioxide; GM-CSF: granulocyte-macrophage colony-stimulating factor.
    Monoclonal Anti Human Lamin B1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology goat anti human lamin b1 antibodies
    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and <t>Lamin</t> B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.
    Goat Anti Human Lamin B1 Antibodies, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology goat anti human lamin b1 polyclonal antibody c 20
    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and <t>Lamin</t> B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.
    Goat Anti Human Lamin B1 Polyclonal Antibody C 20, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    OriGene human lamin b1
    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and <t>Lamin</t> B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.
    Human Lamin B1, supplied by OriGene, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology human lamin b1
    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and <t>Lamin</t> B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.
    Human Lamin B1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher human lamin b1
    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and <t>Lamin</t> B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.
    Human Lamin B1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Genecopoeia human n terminal mcherry tagged lamin b1 preceiver m55
    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and <t>Lamin</t> B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.
    Human N Terminal Mcherry Tagged Lamin B1 Preceiver M55, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc rabbit anti human monoclonal anti lamin b1
    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and <t>Lamin</t> B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.
    Rabbit Anti Human Monoclonal Anti Lamin B1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology rabbit anti human monoclonal lamin b1
    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and <t>Lamin</t> B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.
    Rabbit Anti Human Monoclonal Lamin B1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam rabbit anti human lamin b1 antibody
    Representative western blots of mitochondrial and nuclear proteins in placental micro- and nano- vesicles. Total protein from micro- and nano- vesicles from first trimester human placentae was extracted and the presence of complex IV, <t>lamin</t> B and β-actin was interrogated by western blotting (n = 6 placentae). Placental lysates were included as a positive control.
    Rabbit Anti Human Lamin B1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology rabbit anti human lamin b1 antibody
    Representative western blots of mitochondrial and nuclear proteins in placental micro- and nano- vesicles. Total protein from micro- and nano- vesicles from first trimester human placentae was extracted and the presence of complex IV, <t>lamin</t> B and β-actin was interrogated by western blotting (n = 6 placentae). Placental lysates were included as a positive control.
    Rabbit Anti Human Lamin B1 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rabbit polyclonal anti human lamin b1
    Representative western blots of mitochondrial and nuclear proteins in placental micro- and nano- vesicles. Total protein from micro- and nano- vesicles from first trimester human placentae was extracted and the presence of complex IV, <t>lamin</t> B and β-actin was interrogated by western blotting (n = 6 placentae). Placental lysates were included as a positive control.
    Rabbit Polyclonal Anti Human Lamin B1, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Abcam mouse anti human lamin b1 antibody
    p53 mediated UVB-induced TM downregulation. (A) HaCaT cells (upper panel) or primary human epidermal keratinocytes (lower panel) were UVB irradiated (3 mJ/cm 2 ) at the indicated time points, and cytosolic and nuclear fractions were harvested. p53 and p65 protein levels were determined by Western blotting. α-tubulin or <t>lamin</t> B1 levels were used as cytoplasmic or nuclear loading controls, respectively. (B) Nuclear extracts from UVB-irradiated HaCaT cells (left panel) or human epidermal keratinocytes (right panel) were incubated with proximal TM promoter DNA linked to Dynabeads. Protein-DNA complexes were allowed to form under native conditions and precipitated magnetically. Bound proteins were washed, eluted, and p53 and p65 levels were determined by Western blotting (C) HaCaT cells were UVB irradiated (3 mJ/cm 2 ) for 3 h, and chromatin immunoprecipitation was performed. Chromatin was immunoprecipitated with anti-p53 or anti-p65 antibodies followed by amplification of TM promoter elements by PCR using specific primers. To verify equal loading, 1% of the precipitated chromatin was assayed as input. (D) HaCaT cells were transiently transfected with p53 siRNA or with a GAPDH short hairpin RNA (shRNA) as control. After transfection, cells were exposed to UVB (3 mJ/cm 2 ) for 24 h, and cell lysate was harvested for Western blot analysis to assess p53 and TM levels. α-tubulin was used as a loading control.
    Mouse Anti Human Lamin B1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology polyclonal goat anti human lamin b1
    p53 mediated UVB-induced TM downregulation. (A) HaCaT cells (upper panel) or primary human epidermal keratinocytes (lower panel) were UVB irradiated (3 mJ/cm 2 ) at the indicated time points, and cytosolic and nuclear fractions were harvested. p53 and p65 protein levels were determined by Western blotting. α-tubulin or <t>lamin</t> B1 levels were used as cytoplasmic or nuclear loading controls, respectively. (B) Nuclear extracts from UVB-irradiated HaCaT cells (left panel) or human epidermal keratinocytes (right panel) were incubated with proximal TM promoter DNA linked to Dynabeads. Protein-DNA complexes were allowed to form under native conditions and precipitated magnetically. Bound proteins were washed, eluted, and p53 and p65 levels were determined by Western blotting (C) HaCaT cells were UVB irradiated (3 mJ/cm 2 ) for 3 h, and chromatin immunoprecipitation was performed. Chromatin was immunoprecipitated with anti-p53 or anti-p65 antibodies followed by amplification of TM promoter elements by PCR using specific primers. To verify equal loading, 1% of the precipitated chromatin was assayed as input. (D) HaCaT cells were transiently transfected with p53 siRNA or with a GAPDH short hairpin RNA (shRNA) as control. After transfection, cells were exposed to UVB (3 mJ/cm 2 ) for 24 h, and cell lysate was harvested for Western blot analysis to assess p53 and TM levels. α-tubulin was used as a loading control.
    Polyclonal Goat Anti Human Lamin B1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Curcumin induces the processing of caspase-3 and the cleavage of cytoskeletal proteins. PMNs (10 × 10 6 cells/mL) were incubated for the indicated periods of time with 10 or 50 µM curcumin (Cur10 and Cur50, respectively), and (a) the processing of procaspase-3 (proCASP3) or (b) the cleavage of lamin B1 or vimentin (Vim) were assessed by Western blot as described in the ‘Materials and methods’ section. Results are from one representative experiment out of at least three. Arrowhead indicated the 47-kDa fragment of lamin B 1 observed in HL-60, PLB-985 and human PMNs. 22 SA: spontaneous apoptosis; PMNs: polymorphonuclear neutrophil cells; ATO: arsenic trioxide; GM-CSF: granulocyte-macrophage colony-stimulating factor.

    Journal: SAGE Open Medicine

    Article Title: Mechanisms involved in curcumin-induced human neutrophil apoptosis: Evidence that curcumin activates the endoplasmic reticulum stress-induced cell apoptosis pathway

    doi: 10.1177/2050312113488104

    Figure Lengend Snippet: Curcumin induces the processing of caspase-3 and the cleavage of cytoskeletal proteins. PMNs (10 × 10 6 cells/mL) were incubated for the indicated periods of time with 10 or 50 µM curcumin (Cur10 and Cur50, respectively), and (a) the processing of procaspase-3 (proCASP3) or (b) the cleavage of lamin B1 or vimentin (Vim) were assessed by Western blot as described in the ‘Materials and methods’ section. Results are from one representative experiment out of at least three. Arrowhead indicated the 47-kDa fragment of lamin B 1 observed in HL-60, PLB-985 and human PMNs. 22 SA: spontaneous apoptosis; PMNs: polymorphonuclear neutrophil cells; ATO: arsenic trioxide; GM-CSF: granulocyte-macrophage colony-stimulating factor.

    Article Snippet: Membranes were incubated with monoclonal anti-human lamin B1 (1:1000, clone C-20; Santa Cruz Biotechnology, Inc., Santa Cruz, MA), anti-human vimentin (1:1000, clone V9; Santa Cruz Biotechnology, Inc.), anti-heat shock proteins (HSPs – HSP70 (SPA-810) and HSP90 (SPA-830)), purchased from Stressgen (Ann Arbor, MI, USA), overnight at 4°C.

    Techniques: Incubation, Western Blot

    Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.

    Journal: PLoS ONE

    Article Title: Fusion between Hematopoietic and Epithelial Cells in Adult Human Intestine

    doi: 10.1371/journal.pone.0055572

    Figure Lengend Snippet: Epithelial compartmentalization and sex-karyotyping of intestinal cells. (A) Hematoxylin and Eosin stained intestinal biopsy; epithelial compartment is labeled. (B) Adjacent tissue section to that from panel A stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells and the arrowhead points to a Y-chromosome-positive non-epithelial cell. Inset shows a sub-region stained for cytokeratin (blue); arrows and arrowhead serve as positional references. (C) Enlarged views of cells indicated in panel B by arrows and arrowhead; sex-karyotype is indicated for each. (D) Independent patient sample also stained for X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrows indicate Y-chromosome-positive epithelial cells. Dashed lines in all panels indicate boundaries of epithelial and non-epithelial compartments.

    Article Snippet: Immunofluorescence Staining and Microscopy FISH-processed intestinal tissues were incubated in Blocking Buffer (PBS +10% Donkey Serum, 5% bovine serum albumin +0.3% TritonX-100) for 30 min at 24°C followed by incubation with goat-anti-human Lamin B1 antibodies (1∶200, Santa Cruz Biotechnology, CA) in a humidified chamber at 4°C for 12 h. The Lamin B1 antibody was visualized by incubation with a Cy5-conjugated anti-goat antibody (1∶2000; Jackson, PA) at 24°C for 1 h. Coverslips were mounted with ProLong Gold antifade reagent (Invitrogen, NY).

    Techniques: Staining, Labeling

    Differentiation status of cells with abnormal sex-karyotypes. ( A) FABP2/IFABP expression in a control and (B) transplant patient sample with an example of an XXY cell. Brackets indicate differentiated (high Fabp2/Ifabp expression, black brackets) and undifferentiated (low Fabp2/Ifabp expression, gray brackets) regions of epithelium within each sample. (C) Enlarged view of boxed region from B , in an adjacent tissue section stained for Lamin B1.(D) Enlarged view of boxed region from panel C, showing X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrowhead indicates the same nucleus in panels C and D. Dashed lines indicate boundaries of epithelial and non-epithelial compartments.

    Journal: PLoS ONE

    Article Title: Fusion between Hematopoietic and Epithelial Cells in Adult Human Intestine

    doi: 10.1371/journal.pone.0055572

    Figure Lengend Snippet: Differentiation status of cells with abnormal sex-karyotypes. ( A) FABP2/IFABP expression in a control and (B) transplant patient sample with an example of an XXY cell. Brackets indicate differentiated (high Fabp2/Ifabp expression, black brackets) and undifferentiated (low Fabp2/Ifabp expression, gray brackets) regions of epithelium within each sample. (C) Enlarged view of boxed region from B , in an adjacent tissue section stained for Lamin B1.(D) Enlarged view of boxed region from panel C, showing X- (green) and Y- (red) chromosomes and Lamin B1 (white). Arrowhead indicates the same nucleus in panels C and D. Dashed lines indicate boundaries of epithelial and non-epithelial compartments.

    Article Snippet: Immunofluorescence Staining and Microscopy FISH-processed intestinal tissues were incubated in Blocking Buffer (PBS +10% Donkey Serum, 5% bovine serum albumin +0.3% TritonX-100) for 30 min at 24°C followed by incubation with goat-anti-human Lamin B1 antibodies (1∶200, Santa Cruz Biotechnology, CA) in a humidified chamber at 4°C for 12 h. The Lamin B1 antibody was visualized by incubation with a Cy5-conjugated anti-goat antibody (1∶2000; Jackson, PA) at 24°C for 1 h. Coverslips were mounted with ProLong Gold antifade reagent (Invitrogen, NY).

    Techniques: Expressing, Staining

    Representative western blots of mitochondrial and nuclear proteins in placental micro- and nano- vesicles. Total protein from micro- and nano- vesicles from first trimester human placentae was extracted and the presence of complex IV, lamin B and β-actin was interrogated by western blotting (n = 6 placentae). Placental lysates were included as a positive control.

    Journal: Scientific Reports

    Article Title: Antiphospholipid antibodies increase the levels of mitochondrial DNA in placental extracellular vesicles: Alarmin-g for preeclampsia

    doi: 10.1038/s41598-017-16448-5

    Figure Lengend Snippet: Representative western blots of mitochondrial and nuclear proteins in placental micro- and nano- vesicles. Total protein from micro- and nano- vesicles from first trimester human placentae was extracted and the presence of complex IV, lamin B and β-actin was interrogated by western blotting (n = 6 placentae). Placental lysates were included as a positive control.

    Article Snippet: Membranes were then probed with rabbit anti-human complex IV (Thermo Fisher Scientific, A-6404, 1:750) or rabbit anti-human lamin B (Abcam, ab16048, 1:500) antibodies before applying HRP-conjugated anti-rabbit IgG antibodies (Jackson ImmunoResearch, 1:2000).

    Techniques: Western Blot, Positive Control

    p53 mediated UVB-induced TM downregulation. (A) HaCaT cells (upper panel) or primary human epidermal keratinocytes (lower panel) were UVB irradiated (3 mJ/cm 2 ) at the indicated time points, and cytosolic and nuclear fractions were harvested. p53 and p65 protein levels were determined by Western blotting. α-tubulin or lamin B1 levels were used as cytoplasmic or nuclear loading controls, respectively. (B) Nuclear extracts from UVB-irradiated HaCaT cells (left panel) or human epidermal keratinocytes (right panel) were incubated with proximal TM promoter DNA linked to Dynabeads. Protein-DNA complexes were allowed to form under native conditions and precipitated magnetically. Bound proteins were washed, eluted, and p53 and p65 levels were determined by Western blotting (C) HaCaT cells were UVB irradiated (3 mJ/cm 2 ) for 3 h, and chromatin immunoprecipitation was performed. Chromatin was immunoprecipitated with anti-p53 or anti-p65 antibodies followed by amplification of TM promoter elements by PCR using specific primers. To verify equal loading, 1% of the precipitated chromatin was assayed as input. (D) HaCaT cells were transiently transfected with p53 siRNA or with a GAPDH short hairpin RNA (shRNA) as control. After transfection, cells were exposed to UVB (3 mJ/cm 2 ) for 24 h, and cell lysate was harvested for Western blot analysis to assess p53 and TM levels. α-tubulin was used as a loading control.

    Journal: PLoS ONE

    Article Title: UVB Irradiation Regulates ERK1/2- and p53-Dependent Thrombomodulin Expression in Human Keratinocytes

    doi: 10.1371/journal.pone.0067632

    Figure Lengend Snippet: p53 mediated UVB-induced TM downregulation. (A) HaCaT cells (upper panel) or primary human epidermal keratinocytes (lower panel) were UVB irradiated (3 mJ/cm 2 ) at the indicated time points, and cytosolic and nuclear fractions were harvested. p53 and p65 protein levels were determined by Western blotting. α-tubulin or lamin B1 levels were used as cytoplasmic or nuclear loading controls, respectively. (B) Nuclear extracts from UVB-irradiated HaCaT cells (left panel) or human epidermal keratinocytes (right panel) were incubated with proximal TM promoter DNA linked to Dynabeads. Protein-DNA complexes were allowed to form under native conditions and precipitated magnetically. Bound proteins were washed, eluted, and p53 and p65 levels were determined by Western blotting (C) HaCaT cells were UVB irradiated (3 mJ/cm 2 ) for 3 h, and chromatin immunoprecipitation was performed. Chromatin was immunoprecipitated with anti-p53 or anti-p65 antibodies followed by amplification of TM promoter elements by PCR using specific primers. To verify equal loading, 1% of the precipitated chromatin was assayed as input. (D) HaCaT cells were transiently transfected with p53 siRNA or with a GAPDH short hairpin RNA (shRNA) as control. After transfection, cells were exposed to UVB (3 mJ/cm 2 ) for 24 h, and cell lysate was harvested for Western blot analysis to assess p53 and TM levels. α-tubulin was used as a loading control.

    Article Snippet: The membrane was blocked in 5% fat-free milk in PBST buffer (PBS with 0.05% Tween-20) followed by incubation overnight with primary antibodies (mouse anti-human TM antibodies (1∶1000), mouse anti-human pERK antibodies (1∶1000), mouse anti-human ERK antibodies (1∶1000), mouse anti-human α-tubulin antibodies (1∶2000), mouse anti-human GAPDH antibodies (1∶2000) (Santa Cruz Biotech, CA, USA), mouse anti-human p53 antibodies (1∶1000), mouse anti-human NF-κB antibodies (1∶1000), mouse anti-human fibrillarin antibodies (1∶2000), and mouse anti-human lamin B1 antibody (1∶1000) (Abcam, Cambridge, MA) diluted in PBST buffer.

    Techniques: Irradiation, Western Blot, Incubation, Chromatin Immunoprecipitation, Immunoprecipitation, Amplification, Polymerase Chain Reaction, Transfection, shRNA