hsp 70 Stressgen Inc Search Results


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  • 88
    Cell Signaling Technology Inc anti hsp 70
    Anti Hsp 70, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Assay Designs Inc surface hsp70
    Intracellular <t>HSP70</t> during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample
    Surface Hsp70, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 77/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Assay Designs Inc stressgen hsp70 elisa kit
    Intracellular <t>HSP70</t> during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample
    Stressgen Hsp70 Elisa Kit, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 78/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Assay Designs Inc anti hsp70 antibody
    Loss of pY15-CDK5 and insolubilization of total CDK5 in cells exposed to hyperthermia. A , PErTA70 cells that were either not induced <t>(−HSP70)</t> or induced to express HSP70 (+HSP70) were incubated at 42, 43, or 44 °C for 1 h and then returned to 37 °C for 6 h. Control cells remained at 37 °C. Cells were lysed in buffer containing 1% Triton X-100 and following centrifugation the Triton X-100 soluble fraction was examined by SDS-PAGE and Western blotting. B , quantification of the results shown in A (mean ± S.E., n = 3, * indicates a significant difference between −HSP70 and +HSP70 cells p
    Anti Hsp70 Antibody, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 95/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Cell Signaling Technology Inc hsp70
    The centrosome is a substrate organelle for molecular chaperone <t>Hsp70.</t> (See also Supplemental Figures S1–S3.) (A) Left, images of maximum projections from cntrl and HS cells as indicated show that upon HS, Hsp70 accumulates at the centrosome and in the nucleus (inset, arrow points at the centrosome); bar, 10 μm. Semiquantitative analysis of integrated intensity (×10 5 arbitrary units) of centrosomal Hsp70 before and after HS (15–20 centrosomes/sample, mean ± SEM). (B) Superresolution images (OMX) demonstrate loss of γ-tubulin (red) from HS cells and recruitment of Hsp70 (green) as an outer layer in stressed cells; bar, 0.2 μm. (C) Immunoprecipitation (IP) of Hsp70 pulls down PCNT. Immunoglobulin G (IgG), control. (D) Centrosome-targeted Hsp70 (cHsp70) protects γ-tubulin from HS, whereas membrane-targeted Hsp70 (mHsp70) does not. Data are shown as semiquantitative profiles of confocal microscopy images. Right, percentage of cells with centrosomal γ-tubulin after HS in stably expressing centrosome-(cHsp70) or membrane-(mHsp70) targeted Hsp70-expressing cells (four experiments, 500–600 cells/sample, mean ± SD, one-way analysis of variance [ANOVA] combined with Tukey’s multiple comparison test). (E) cHsp70, but not the chaperone-negative mutant cHsp70D10S, protects γ-tubulin signal at the centrosome; semiquantitative analysis of integrated γ-tubulin signal intensity (×10 5 arbitrary units, mean ± SEM). (F) cHsp70 protects centrosomal Centrin2 from HS, whereas mHsp70 does not. Data are shown as semiquantitative profiles of confocal microscopy images. Right, percentage of cells with centrosome-localized Centrin2 after HS in stably expressing cHsp70- or mHsp70-targeted Hsp70 cells (three experiments, 500–600 cells/sample, mean ± SD, one-way ANOVA combined with Tukey’s multiple comparison test).
    Hsp70, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1171 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Assay Designs Inc pe conjugated hsp70 monoclonal antibody
    Intracellular <t>HSP70</t> during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample
    Pe Conjugated Hsp70 Monoclonal Antibody, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioTechniques mouse monoclonal anti hsp70
    Intracellular <t>HSP70</t> during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample
    Mouse Monoclonal Anti Hsp70, supplied by BioTechniques, 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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    77
    Assay Designs Inc antihuman hsp70 elisa kit
    Intracellular <t>HSP70</t> during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample
    Antihuman Hsp70 Elisa Kit, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Assay Designs Inc hsp70 elisa kit
    Intracellular <t>HSP70</t> during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample
    Hsp70 Elisa Kit, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 77/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stressgen Biotechnologies mouse anti hsp70
    Effects of <t>hsp70-CaM</t> interaction on cell cycle and apoptosis. a Time-course observation of living cell that co-expressed CFP-CaM and YFP-Hsp70.The observation began at later G1 stage and the image was recorded every 5 min. CaM was translocated
    Mouse Anti Hsp70, supplied by Stressgen Biotechnologies, used in various techniques. Bioz Stars score: 85/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc mouse anti hsp70
    Anterior horn region of lumbar spinal cord sections from mice at 220 days. Motor neuron choline acetyltransferase (ChAT) positive cells (A-C, arrows) co-localized with SOD1 positive staining (D-F, arrows) . H46R/H48Q mice exhibited small intracellular peri-nuclear SOD1 reactive punctae (E, arrows) , while H46R/H48QxHSF1 tissues had a more even intracellular distribution of SOD1 staining (F) . This corresponded with differences in <t>HSP70</t> distribution to SOD1 reactive punctae in H46R/H48Q mice (H) while in contrast, co-localization with ChAT reactive cells in H46R/H48QxHSF1 mice was stronger (I) . Scale bar represents 10 μm.
    Mouse Anti Hsp70, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Assay Designs Inc anti hsp70 rabbit antibodies
    Treatment with MG132 activates stress. (A) Inhibition of proteasome activated <t>hsp70</t> synthesis. Control HeLa cells, cells treated for 4 h with MG132, and 4 h VSV-infected cells were incubated for last 30 min with S 35 methionine/cysteine. Cytoplasmic proteins were precipitated with anti-hsp70 and anti-P-VSV Abs. Precipitated proteins were analyzed by electrophoresis and autoradiography. (B) MG132 stimulated eIF2α phosphorylation. HeLa cells were treated with 1 µM of thapsigargin for 1 h and with 5 µM of MG132 for 4 h. 10 µg of protein extracts were analyzed with Abs specific for eIF2α and eIF2α- phosphate (eIF2α-P). Hsp70 is a marker of MG132 activated stress. Keratin 18 (K18) is a loading control.
    Anti Hsp70 Rabbit Antibodies, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Assay Designs Inc mouse anti human hsp70 mab
    Treatment with MG132 activates stress. (A) Inhibition of proteasome activated <t>hsp70</t> synthesis. Control HeLa cells, cells treated for 4 h with MG132, and 4 h VSV-infected cells were incubated for last 30 min with S 35 methionine/cysteine. Cytoplasmic proteins were precipitated with anti-hsp70 and anti-P-VSV Abs. Precipitated proteins were analyzed by electrophoresis and autoradiography. (B) MG132 stimulated eIF2α phosphorylation. HeLa cells were treated with 1 µM of thapsigargin for 1 h and with 5 µM of MG132 for 4 h. 10 µg of protein extracts were analyzed with Abs specific for eIF2α and eIF2α- phosphate (eIF2α-P). Hsp70 is a marker of MG132 activated stress. Keratin 18 (K18) is a loading control.
    Mouse Anti Human Hsp70 Mab, supplied by Assay Designs Inc, 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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    Cell Signaling Technology Inc rabbit anti hsc70
    <t>Hsc70</t> and PKCι distribution depend on the integrity of intermediate filaments in small intestine villi of K8 knockout mice. Small intestines were harvested from K8-null mice (C,D,I,J) or from K18 +/– littermates (A,B,E-H), fixed in 10%
    Rabbit Anti Hsc70, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 79/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Intracellular HSP70 during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample

    Journal: Cell Stress & Chaperones

    Article Title: Eleven days of moderate exercise and heat exposure induces acclimation without significant HSP70 and apoptosis responses of lymphocytes in college-aged males

    doi: 10.1007/s12192-011-0283-5

    Figure Lengend Snippet: Intracellular HSP70 during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample

    Article Snippet: Intracellular and surface HSP70 (SPA-820PE, Stressgen, Assay Designs), and annexin V (ab14085, Abcam Inc.), as a marker of early apoptosis, were measured on CD3+ and CD4+ (sc-70624, sc-70670, Santa Cruz Biotechnology) gated lymphocytes.

    Techniques: Flow Cytometry

    HSP70 expression during ex vivo heat shock of lymphocytes. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b Percent HSP70 + cells out of total live lymphocytes, normalized to control, day 1. Error bars represent ± 2SE

    Journal: Cell Stress & Chaperones

    Article Title: Eleven days of moderate exercise and heat exposure induces acclimation without significant HSP70 and apoptosis responses of lymphocytes in college-aged males

    doi: 10.1007/s12192-011-0283-5

    Figure Lengend Snippet: HSP70 expression during ex vivo heat shock of lymphocytes. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b Percent HSP70 + cells out of total live lymphocytes, normalized to control, day 1. Error bars represent ± 2SE

    Article Snippet: Intracellular and surface HSP70 (SPA-820PE, Stressgen, Assay Designs), and annexin V (ab14085, Abcam Inc.), as a marker of early apoptosis, were measured on CD3+ and CD4+ (sc-70624, sc-70670, Santa Cruz Biotechnology) gated lymphocytes.

    Techniques: Expressing, Ex Vivo

    Loss of pY15-CDK5 and insolubilization of total CDK5 in cells exposed to hyperthermia. A , PErTA70 cells that were either not induced (−HSP70) or induced to express HSP70 (+HSP70) were incubated at 42, 43, or 44 °C for 1 h and then returned to 37 °C for 6 h. Control cells remained at 37 °C. Cells were lysed in buffer containing 1% Triton X-100 and following centrifugation the Triton X-100 soluble fraction was examined by SDS-PAGE and Western blotting. B , quantification of the results shown in A (mean ± S.E., n = 3, * indicates a significant difference between −HSP70 and +HSP70 cells p

    Journal: The Journal of Biological Chemistry

    Article Title: Heat Shock Inhibition of CDK5 Increases NOXA Levels through miR-23a Repression *

    doi: 10.1074/jbc.M114.625988

    Figure Lengend Snippet: Loss of pY15-CDK5 and insolubilization of total CDK5 in cells exposed to hyperthermia. A , PErTA70 cells that were either not induced (−HSP70) or induced to express HSP70 (+HSP70) were incubated at 42, 43, or 44 °C for 1 h and then returned to 37 °C for 6 h. Control cells remained at 37 °C. Cells were lysed in buffer containing 1% Triton X-100 and following centrifugation the Triton X-100 soluble fraction was examined by SDS-PAGE and Western blotting. B , quantification of the results shown in A (mean ± S.E., n = 3, * indicates a significant difference between −HSP70 and +HSP70 cells p

    Article Snippet: The following antibodies were used for immunoblotting: Actin (ACTN05: NeoMarkers, Fremont, CA), CDK5 (2506: Cell Signaling Technology, Danvers, MA), cleaved caspase-3 Asp175 (9664: Cell Signaling Technology), cytochrome c (65981A: BD Biosciences PharMingen, Mississauga, Ontario, Canada), c-myc from 9E10 hybridoma supernatant, HSP60 (SPC-105: StressMarq Biosciences, Victoria, British Columbia, Canada), HSP70 (C92F3A-5: Stressgen/Assay Designs, Ann Arbor, MI, USA), HtrA2 (AF1458: R & D Systems/Cedarlane, Burlington, Ontario, Canada), MCL1 (SC-819: Santa Cruz Biotechnology, Santa Cruz, CA), NOXA (ALX-804–408: Enzo Life Sciences), p35/25 (2680: Cell Signaling Technology), phospho-MAPK/CDK substrates (PXS*P or S*PXR/K, 2325: Cell Signaling Technology), phospho-CDK5 Tyr15 (CG1085: Cell Applications, San Diego, CA), tubulin (MABT205: Millipore, Billerica, MA).

    Techniques: Incubation, Centrifugation, SDS Page, Western Blot

    The centrosome is a substrate organelle for molecular chaperone Hsp70. (See also Supplemental Figures S1–S3.) (A) Left, images of maximum projections from cntrl and HS cells as indicated show that upon HS, Hsp70 accumulates at the centrosome and in the nucleus (inset, arrow points at the centrosome); bar, 10 μm. Semiquantitative analysis of integrated intensity (×10 5 arbitrary units) of centrosomal Hsp70 before and after HS (15–20 centrosomes/sample, mean ± SEM). (B) Superresolution images (OMX) demonstrate loss of γ-tubulin (red) from HS cells and recruitment of Hsp70 (green) as an outer layer in stressed cells; bar, 0.2 μm. (C) Immunoprecipitation (IP) of Hsp70 pulls down PCNT. Immunoglobulin G (IgG), control. (D) Centrosome-targeted Hsp70 (cHsp70) protects γ-tubulin from HS, whereas membrane-targeted Hsp70 (mHsp70) does not. Data are shown as semiquantitative profiles of confocal microscopy images. Right, percentage of cells with centrosomal γ-tubulin after HS in stably expressing centrosome-(cHsp70) or membrane-(mHsp70) targeted Hsp70-expressing cells (four experiments, 500–600 cells/sample, mean ± SD, one-way analysis of variance [ANOVA] combined with Tukey’s multiple comparison test). (E) cHsp70, but not the chaperone-negative mutant cHsp70D10S, protects γ-tubulin signal at the centrosome; semiquantitative analysis of integrated γ-tubulin signal intensity (×10 5 arbitrary units, mean ± SEM). (F) cHsp70 protects centrosomal Centrin2 from HS, whereas mHsp70 does not. Data are shown as semiquantitative profiles of confocal microscopy images. Right, percentage of cells with centrosome-localized Centrin2 after HS in stably expressing cHsp70- or mHsp70-targeted Hsp70 cells (three experiments, 500–600 cells/sample, mean ± SD, one-way ANOVA combined with Tukey’s multiple comparison test).

    Journal: Molecular Biology of the Cell

    Article Title: Centrosome-intrinsic mechanisms modulate centrosome integrity during fever

    doi: 10.1091/mbc.E15-03-0158

    Figure Lengend Snippet: The centrosome is a substrate organelle for molecular chaperone Hsp70. (See also Supplemental Figures S1–S3.) (A) Left, images of maximum projections from cntrl and HS cells as indicated show that upon HS, Hsp70 accumulates at the centrosome and in the nucleus (inset, arrow points at the centrosome); bar, 10 μm. Semiquantitative analysis of integrated intensity (×10 5 arbitrary units) of centrosomal Hsp70 before and after HS (15–20 centrosomes/sample, mean ± SEM). (B) Superresolution images (OMX) demonstrate loss of γ-tubulin (red) from HS cells and recruitment of Hsp70 (green) as an outer layer in stressed cells; bar, 0.2 μm. (C) Immunoprecipitation (IP) of Hsp70 pulls down PCNT. Immunoglobulin G (IgG), control. (D) Centrosome-targeted Hsp70 (cHsp70) protects γ-tubulin from HS, whereas membrane-targeted Hsp70 (mHsp70) does not. Data are shown as semiquantitative profiles of confocal microscopy images. Right, percentage of cells with centrosomal γ-tubulin after HS in stably expressing centrosome-(cHsp70) or membrane-(mHsp70) targeted Hsp70-expressing cells (four experiments, 500–600 cells/sample, mean ± SD, one-way analysis of variance [ANOVA] combined with Tukey’s multiple comparison test). (E) cHsp70, but not the chaperone-negative mutant cHsp70D10S, protects γ-tubulin signal at the centrosome; semiquantitative analysis of integrated γ-tubulin signal intensity (×10 5 arbitrary units, mean ± SEM). (F) cHsp70 protects centrosomal Centrin2 from HS, whereas mHsp70 does not. Data are shown as semiquantitative profiles of confocal microscopy images. Right, percentage of cells with centrosome-localized Centrin2 after HS in stably expressing cHsp70- or mHsp70-targeted Hsp70 cells (three experiments, 500–600 cells/sample, mean ± SD, one-way ANOVA combined with Tukey’s multiple comparison test).

    Article Snippet: The following commercial antibodies were used: ninein (ab4447; Abcam [Cambridge, MA]), pericentrin (ab4448; Abcam), cenexin/ODF2 (ab43840; Abcam), centrobin (ab70448; Abcam), Hsp70 (SPA-810; Stressgen [Plymouth Meeting, PA]), PCM1 (5213; Cell Signaling [Danvers, MA]), glyceraldehyde-3-phosphate dehydrogenase (sc-32233; Santa Cruz Biotechnology [Dallas, TX]), rootletin (sc-67824; Santa Cruz Biotechnology), SAS-6 (sc-81431; Santa Cruz Biotechnology), Cep83 (HPA038161; Atlas [Stockholm, Sweden]), Septin7 (sc-20620; Santa Cruz Biotechnology), MLKP (sc-867; Santa Cruz Biotechnology), ubiquitin (550944; BD Transduction [San Jose, CA]), p150 (P50520-050; BD Biosciences), RacGAP (ab2270; Abcam), acetylated tubulin (T 6793; Sigma-Aldrich [St. Lous, MO]), EB1 (610534; BD Transduction), CREST (15-234-0001; Antibodies Incorporated [Davis, CA]), CenpE (ab5093; Abcam), ATP5A (ab14748; Abcam), mitochondrial Hsp70 (MA3-028; Affinity Bioreagents [Golden, CO]), GFP (DSHB-GFP-12A6; DSHB Biology [Iowa City, IA]), and GFP (sc-8334; Santa Cruz Biotechnology).

    Techniques: Immunoprecipitation, Confocal Microscopy, Stable Transfection, Expressing, Mutagenesis

    Centrosome degradation is specific. (A) Mitochondria are not degraded during HS. Maximum projections from confocal microscopy images show representative images of cntrl and HS cells immunostained with ATP5A (left; bar,10 μm) and mitochondrial Hsp70 (green) and γ- tubulin (red); middle and right, inset, centrosomes (bar, 10 μm). (B) Kinetochores are not degraded during HS. Maximum projections from confocal microscopy images show representative images of prometaphase cells from cntrl and HS cells immunostained with CenpE (green) and CREST (red); bar 10 μm. (C) Semiquantitative analysis of integrated intensity of the midbody marker RacGap (×10 5 arbitrary units, mean ± SEM) demonstrates no significant difference in signal intensity in HS cells. (D) Semiquantitative analysis of integrated intensity of the midbody marker mitotic kinesin-like protein (×10 5 arbitrary units, mean ± SEM) demonstrates no significant difference in signal intensity in HS cells. (E) Maximum projections from confocal microscopy images show representative images of cntrl and HS cells immunostained with Sept7 (red, decorates midbody, MB, and centrosomes, Cen) and Centrin2 (green, as centrosome marker); bar, 5 μm. HS results in loss of Septin7 from centrosomes but not from midbodies in the same cell. (F) Disruption of Septin7 signal after HS is shown at the centrosome but not at the midbody by semiquantitative analysis of integrated intensity (×10 5 arbitrary units, mean ± SEM).

    Journal: Molecular Biology of the Cell

    Article Title: Centrosome-intrinsic mechanisms modulate centrosome integrity during fever

    doi: 10.1091/mbc.E15-03-0158

    Figure Lengend Snippet: Centrosome degradation is specific. (A) Mitochondria are not degraded during HS. Maximum projections from confocal microscopy images show representative images of cntrl and HS cells immunostained with ATP5A (left; bar,10 μm) and mitochondrial Hsp70 (green) and γ- tubulin (red); middle and right, inset, centrosomes (bar, 10 μm). (B) Kinetochores are not degraded during HS. Maximum projections from confocal microscopy images show representative images of prometaphase cells from cntrl and HS cells immunostained with CenpE (green) and CREST (red); bar 10 μm. (C) Semiquantitative analysis of integrated intensity of the midbody marker RacGap (×10 5 arbitrary units, mean ± SEM) demonstrates no significant difference in signal intensity in HS cells. (D) Semiquantitative analysis of integrated intensity of the midbody marker mitotic kinesin-like protein (×10 5 arbitrary units, mean ± SEM) demonstrates no significant difference in signal intensity in HS cells. (E) Maximum projections from confocal microscopy images show representative images of cntrl and HS cells immunostained with Sept7 (red, decorates midbody, MB, and centrosomes, Cen) and Centrin2 (green, as centrosome marker); bar, 5 μm. HS results in loss of Septin7 from centrosomes but not from midbodies in the same cell. (F) Disruption of Septin7 signal after HS is shown at the centrosome but not at the midbody by semiquantitative analysis of integrated intensity (×10 5 arbitrary units, mean ± SEM).

    Article Snippet: The following commercial antibodies were used: ninein (ab4447; Abcam [Cambridge, MA]), pericentrin (ab4448; Abcam), cenexin/ODF2 (ab43840; Abcam), centrobin (ab70448; Abcam), Hsp70 (SPA-810; Stressgen [Plymouth Meeting, PA]), PCM1 (5213; Cell Signaling [Danvers, MA]), glyceraldehyde-3-phosphate dehydrogenase (sc-32233; Santa Cruz Biotechnology [Dallas, TX]), rootletin (sc-67824; Santa Cruz Biotechnology), SAS-6 (sc-81431; Santa Cruz Biotechnology), Cep83 (HPA038161; Atlas [Stockholm, Sweden]), Septin7 (sc-20620; Santa Cruz Biotechnology), MLKP (sc-867; Santa Cruz Biotechnology), ubiquitin (550944; BD Transduction [San Jose, CA]), p150 (P50520-050; BD Biosciences), RacGAP (ab2270; Abcam), acetylated tubulin (T 6793; Sigma-Aldrich [St. Lous, MO]), EB1 (610534; BD Transduction), CREST (15-234-0001; Antibodies Incorporated [Davis, CA]), CenpE (ab5093; Abcam), ATP5A (ab14748; Abcam), mitochondrial Hsp70 (MA3-028; Affinity Bioreagents [Golden, CO]), GFP (DSHB-GFP-12A6; DSHB Biology [Iowa City, IA]), and GFP (sc-8334; Santa Cruz Biotechnology).

    Techniques: Confocal Microscopy, Marker

    Hsp70 enables centrosome recovery, and protects centrosome functions after HS. (A) Semiquantitative analysis (×10 5 arbitrary units) of Centrin2 signal intensity shows centrosome disruption and recovery after HS during 24 h. (B) Semiquantitative intensity profiles of PCNT from cells recovered for 24 h at 37˚C after HS demonstrate that Hsp70 depletion impairs centrosome recovery. (C) Confocal microscopy images demonstrating rescue of MT regrowth early after microtubule depolymerization (α-tubulin, 1 min of regrowth) after HS in cells expressing the centrosome targeting protein cHsp70 but not the membrane-targeting protein mHsp70. Percentage of the cells positive for detectable MT regrowth 1 min after HS exposure in RPE cell lines expressing cHsp70 or mHsp70 (five experiments, 400–500 cells/sample, mean ± SD). (D) Percentage of the cells with cilia after HS in cells expressing cHsp70 or mHsp70 (three experiments, 400–500 cells/sample, mean ± SD). Maximum projections of ciliated cells (cilia marker, glutamylated tubulin, red; centrosome marker, γ-tubulin, green) after HS exposure in cells expressing cHsp70 or mHsp70. (E) Centrosomal Hsp70 protects centrosome from HS-induced damage in IS conjugates; left, ×10 5 arbitrary units, 10–40 centrosomes/sample, mean ±S EM; middle, distance between centrosome and IS, micrometers. Right, γ-tubulin, red, over DIC images; bar, 5 μm.

    Journal: Molecular Biology of the Cell

    Article Title: Centrosome-intrinsic mechanisms modulate centrosome integrity during fever

    doi: 10.1091/mbc.E15-03-0158

    Figure Lengend Snippet: Hsp70 enables centrosome recovery, and protects centrosome functions after HS. (A) Semiquantitative analysis (×10 5 arbitrary units) of Centrin2 signal intensity shows centrosome disruption and recovery after HS during 24 h. (B) Semiquantitative intensity profiles of PCNT from cells recovered for 24 h at 37˚C after HS demonstrate that Hsp70 depletion impairs centrosome recovery. (C) Confocal microscopy images demonstrating rescue of MT regrowth early after microtubule depolymerization (α-tubulin, 1 min of regrowth) after HS in cells expressing the centrosome targeting protein cHsp70 but not the membrane-targeting protein mHsp70. Percentage of the cells positive for detectable MT regrowth 1 min after HS exposure in RPE cell lines expressing cHsp70 or mHsp70 (five experiments, 400–500 cells/sample, mean ± SD). (D) Percentage of the cells with cilia after HS in cells expressing cHsp70 or mHsp70 (three experiments, 400–500 cells/sample, mean ± SD). Maximum projections of ciliated cells (cilia marker, glutamylated tubulin, red; centrosome marker, γ-tubulin, green) after HS exposure in cells expressing cHsp70 or mHsp70. (E) Centrosomal Hsp70 protects centrosome from HS-induced damage in IS conjugates; left, ×10 5 arbitrary units, 10–40 centrosomes/sample, mean ±S EM; middle, distance between centrosome and IS, micrometers. Right, γ-tubulin, red, over DIC images; bar, 5 μm.

    Article Snippet: The following commercial antibodies were used: ninein (ab4447; Abcam [Cambridge, MA]), pericentrin (ab4448; Abcam), cenexin/ODF2 (ab43840; Abcam), centrobin (ab70448; Abcam), Hsp70 (SPA-810; Stressgen [Plymouth Meeting, PA]), PCM1 (5213; Cell Signaling [Danvers, MA]), glyceraldehyde-3-phosphate dehydrogenase (sc-32233; Santa Cruz Biotechnology [Dallas, TX]), rootletin (sc-67824; Santa Cruz Biotechnology), SAS-6 (sc-81431; Santa Cruz Biotechnology), Cep83 (HPA038161; Atlas [Stockholm, Sweden]), Septin7 (sc-20620; Santa Cruz Biotechnology), MLKP (sc-867; Santa Cruz Biotechnology), ubiquitin (550944; BD Transduction [San Jose, CA]), p150 (P50520-050; BD Biosciences), RacGAP (ab2270; Abcam), acetylated tubulin (T 6793; Sigma-Aldrich [St. Lous, MO]), EB1 (610534; BD Transduction), CREST (15-234-0001; Antibodies Incorporated [Davis, CA]), CenpE (ab5093; Abcam), ATP5A (ab14748; Abcam), mitochondrial Hsp70 (MA3-028; Affinity Bioreagents [Golden, CO]), GFP (DSHB-GFP-12A6; DSHB Biology [Iowa City, IA]), and GFP (sc-8334; Santa Cruz Biotechnology).

    Techniques: Confocal Microscopy, Expressing, Marker

    Intracellular HSP70 during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample

    Journal: Cell Stress & Chaperones

    Article Title: Eleven days of moderate exercise and heat exposure induces acclimation without significant HSP70 and apoptosis responses of lymphocytes in college-aged males

    doi: 10.1007/s12192-011-0283-5

    Figure Lengend Snippet: Intracellular HSP70 during acclimation. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b percent HSP70 + cells out of total live lymphocytes, normalized to control day 1; c representative flow plots of each day of acclimation for one sample

    Article Snippet: After two washes (centrifugation at 2,500 rpm, 5 min), cells were stained with 10 μL of pre-diluted PE-conjugated HSP70 monoclonal antibody (SPA-820PE, Stressgen, Assay Designs, Ann Arbor, MI) (1 μL + 9 μL PBS) and allowed to incubate on ice in the dark for 30 min.

    Techniques: Flow Cytometry

    HSP70 expression during ex vivo heat shock of lymphocytes. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b Percent HSP70 + cells out of total live lymphocytes, normalized to control, day 1. Error bars represent ± 2SE

    Journal: Cell Stress & Chaperones

    Article Title: Eleven days of moderate exercise and heat exposure induces acclimation without significant HSP70 and apoptosis responses of lymphocytes in college-aged males

    doi: 10.1007/s12192-011-0283-5

    Figure Lengend Snippet: HSP70 expression during ex vivo heat shock of lymphocytes. a MFI of HSP70 + lymphocytes, normalized to control, day 1; b Percent HSP70 + cells out of total live lymphocytes, normalized to control, day 1. Error bars represent ± 2SE

    Article Snippet: After two washes (centrifugation at 2,500 rpm, 5 min), cells were stained with 10 μL of pre-diluted PE-conjugated HSP70 monoclonal antibody (SPA-820PE, Stressgen, Assay Designs, Ann Arbor, MI) (1 μL + 9 μL PBS) and allowed to incubate on ice in the dark for 30 min.

    Techniques: Expressing, Ex Vivo

    Effects of hsp70-CaM interaction on cell cycle and apoptosis. a Time-course observation of living cell that co-expressed CFP-CaM and YFP-Hsp70.The observation began at later G1 stage and the image was recorded every 5 min. CaM was translocated

    Journal:

    Article Title: The association of CaM and Hsp70 regulates S-phase arrest and apoptosis in a spatially and temporally dependent manner in human cells

    doi: 10.1007/s12192-008-0088-3

    Figure Lengend Snippet: Effects of hsp70-CaM interaction on cell cycle and apoptosis. a Time-course observation of living cell that co-expressed CFP-CaM and YFP-Hsp70.The observation began at later G1 stage and the image was recorded every 5 min. CaM was translocated

    Article Snippet: The membranes were then incubated with primary mouse anti-laminB2 (1:500 dilution, Santa Cruz, CA, USA), mouse anti-HSP70 (1:500 dilution, Stressgen Biotechnologies, Inc., San Diego, CA) and mouse anti-HSP90 (SC-69703; 1:500 dilution, Santa Cruz, CA, USA) antibodies.

    Techniques: Chick Chorioallantoic Membrane Assay

    Immunoprecipitation of CaM with Hsp90, Hsp70 and Lamin B2. All selected proteins are shown to interact with CaM in vitro. a CaM and Lamin B2; b CaM and Hsp90; c CaM can associate with Hsp70 in both HCC7402 and ECV304 cells, and its binding is increased

    Journal:

    Article Title: The association of CaM and Hsp70 regulates S-phase arrest and apoptosis in a spatially and temporally dependent manner in human cells

    doi: 10.1007/s12192-008-0088-3

    Figure Lengend Snippet: Immunoprecipitation of CaM with Hsp90, Hsp70 and Lamin B2. All selected proteins are shown to interact with CaM in vitro. a CaM and Lamin B2; b CaM and Hsp90; c CaM can associate with Hsp70 in both HCC7402 and ECV304 cells, and its binding is increased

    Article Snippet: The membranes were then incubated with primary mouse anti-laminB2 (1:500 dilution, Santa Cruz, CA, USA), mouse anti-HSP70 (1:500 dilution, Stressgen Biotechnologies, Inc., San Diego, CA) and mouse anti-HSP90 (SC-69703; 1:500 dilution, Santa Cruz, CA, USA) antibodies.

    Techniques: Immunoprecipitation, Chick Chorioallantoic Membrane Assay, In Vitro, Binding Assay

    Hsp70 associated with CaM in a temporally and spatially dependent manner. a IP result of Hsp70 and CaM showed that the Hsp70 and CaM complex was found to be more abundant in the nucleus in synchronized S-phase cells but not in total interphase cells.

    Journal:

    Article Title: The association of CaM and Hsp70 regulates S-phase arrest and apoptosis in a spatially and temporally dependent manner in human cells

    doi: 10.1007/s12192-008-0088-3

    Figure Lengend Snippet: Hsp70 associated with CaM in a temporally and spatially dependent manner. a IP result of Hsp70 and CaM showed that the Hsp70 and CaM complex was found to be more abundant in the nucleus in synchronized S-phase cells but not in total interphase cells.

    Article Snippet: The membranes were then incubated with primary mouse anti-laminB2 (1:500 dilution, Santa Cruz, CA, USA), mouse anti-HSP70 (1:500 dilution, Stressgen Biotechnologies, Inc., San Diego, CA) and mouse anti-HSP90 (SC-69703; 1:500 dilution, Santa Cruz, CA, USA) antibodies.

    Techniques: Chick Chorioallantoic Membrane Assay

    Immunofluorescence staining of CaM with Hsp70, LaminB2 or Actin to show their co-localization. a cytosol co-localization of CaM ( red ) and Hsp70 ( green ); b nuclear co-localization of CaM ( red ) and Hsp70 ( green ); c nuclear membrane co-localization of CaM

    Journal:

    Article Title: The association of CaM and Hsp70 regulates S-phase arrest and apoptosis in a spatially and temporally dependent manner in human cells

    doi: 10.1007/s12192-008-0088-3

    Figure Lengend Snippet: Immunofluorescence staining of CaM with Hsp70, LaminB2 or Actin to show their co-localization. a cytosol co-localization of CaM ( red ) and Hsp70 ( green ); b nuclear co-localization of CaM ( red ) and Hsp70 ( green ); c nuclear membrane co-localization of CaM

    Article Snippet: The membranes were then incubated with primary mouse anti-laminB2 (1:500 dilution, Santa Cruz, CA, USA), mouse anti-HSP70 (1:500 dilution, Stressgen Biotechnologies, Inc., San Diego, CA) and mouse anti-HSP90 (SC-69703; 1:500 dilution, Santa Cruz, CA, USA) antibodies.

    Techniques: Immunofluorescence, Staining, Chick Chorioallantoic Membrane Assay

    Anterior horn region of lumbar spinal cord sections from mice at 220 days. Motor neuron choline acetyltransferase (ChAT) positive cells (A-C, arrows) co-localized with SOD1 positive staining (D-F, arrows) . H46R/H48Q mice exhibited small intracellular peri-nuclear SOD1 reactive punctae (E, arrows) , while H46R/H48QxHSF1 tissues had a more even intracellular distribution of SOD1 staining (F) . This corresponded with differences in HSP70 distribution to SOD1 reactive punctae in H46R/H48Q mice (H) while in contrast, co-localization with ChAT reactive cells in H46R/H48QxHSF1 mice was stronger (I) . Scale bar represents 10 μm.

    Journal: Molecular Neurodegeneration

    Article Title: Heat shock factor 1 over-expression protects against exposure of hydrophobic residues on mutant SOD1 and early mortality in a mouse model of amyotrophic lateral sclerosis

    doi: 10.1186/1750-1326-8-43

    Figure Lengend Snippet: Anterior horn region of lumbar spinal cord sections from mice at 220 days. Motor neuron choline acetyltransferase (ChAT) positive cells (A-C, arrows) co-localized with SOD1 positive staining (D-F, arrows) . H46R/H48Q mice exhibited small intracellular peri-nuclear SOD1 reactive punctae (E, arrows) , while H46R/H48QxHSF1 tissues had a more even intracellular distribution of SOD1 staining (F) . This corresponded with differences in HSP70 distribution to SOD1 reactive punctae in H46R/H48Q mice (H) while in contrast, co-localization with ChAT reactive cells in H46R/H48QxHSF1 mice was stronger (I) . Scale bar represents 10 μm.

    Article Snippet: Membranes were incubated with sheep anti-human SOD1 (1:1000, Calbiochem), mouse anti-UCHL1 (1:5,000 Abnova#H00007345-M01), mouse anti-Hsp70 (1:10,000, Stressgen; SPA-810), rabbit anti-HSF1 (1:10,000 dilution, Stressgen;SPA-901), heat shock cognate 70 (Hsc70) (1:20,000 dilution, Stressgen SPA-815), rabbit anti-LC3 (1:2,500 dilution, Cell Signaling#2775), rabbit anti-CHIP (1:1000, Cell Signaling#2080), rabbit anti-αB-crystallin (1:2000, Abcam #ab13497), overnight at 4°C with gentle agitation.

    Techniques: Mouse Assay, Staining

    Expression of HSF1 and distribution of HSPs in spinal cord. A) Western blot of HSF1 in spinal cord homogenates from normal and symptomatic control mice normalized with HSC70, demonstrating the over-expression of HSF1 in H46R/H48QxHSF1 mice compared to H46R/H48Q littermates. B) Spinal cords were extracted with detergents of increasing ionic strength as described in the methods. S1 and P1-3 extracts were immunoblotted for inducible HSP70 (B) or αB-crystallin (C) . Asterisks are given at the position of the expected monomeric protein. p is indicated by brackets, bars represent the mean of 6 mice +/- SD.

    Journal: Molecular Neurodegeneration

    Article Title: Heat shock factor 1 over-expression protects against exposure of hydrophobic residues on mutant SOD1 and early mortality in a mouse model of amyotrophic lateral sclerosis

    doi: 10.1186/1750-1326-8-43

    Figure Lengend Snippet: Expression of HSF1 and distribution of HSPs in spinal cord. A) Western blot of HSF1 in spinal cord homogenates from normal and symptomatic control mice normalized with HSC70, demonstrating the over-expression of HSF1 in H46R/H48QxHSF1 mice compared to H46R/H48Q littermates. B) Spinal cords were extracted with detergents of increasing ionic strength as described in the methods. S1 and P1-3 extracts were immunoblotted for inducible HSP70 (B) or αB-crystallin (C) . Asterisks are given at the position of the expected monomeric protein. p is indicated by brackets, bars represent the mean of 6 mice +/- SD.

    Article Snippet: Membranes were incubated with sheep anti-human SOD1 (1:1000, Calbiochem), mouse anti-UCHL1 (1:5,000 Abnova#H00007345-M01), mouse anti-Hsp70 (1:10,000, Stressgen; SPA-810), rabbit anti-HSF1 (1:10,000 dilution, Stressgen;SPA-901), heat shock cognate 70 (Hsc70) (1:20,000 dilution, Stressgen SPA-815), rabbit anti-LC3 (1:2,500 dilution, Cell Signaling#2775), rabbit anti-CHIP (1:1000, Cell Signaling#2080), rabbit anti-αB-crystallin (1:2000, Abcam #ab13497), overnight at 4°C with gentle agitation.

    Techniques: Expressing, Western Blot, Mouse Assay, Over Expression

    Treatment with MG132 activates stress. (A) Inhibition of proteasome activated hsp70 synthesis. Control HeLa cells, cells treated for 4 h with MG132, and 4 h VSV-infected cells were incubated for last 30 min with S 35 methionine/cysteine. Cytoplasmic proteins were precipitated with anti-hsp70 and anti-P-VSV Abs. Precipitated proteins were analyzed by electrophoresis and autoradiography. (B) MG132 stimulated eIF2α phosphorylation. HeLa cells were treated with 1 µM of thapsigargin for 1 h and with 5 µM of MG132 for 4 h. 10 µg of protein extracts were analyzed with Abs specific for eIF2α and eIF2α- phosphate (eIF2α-P). Hsp70 is a marker of MG132 activated stress. Keratin 18 (K18) is a loading control.

    Journal: PLoS ONE

    Article Title: Different Effect of Proteasome Inhibition on Vesicular Stomatitis Virus and Poliovirus Replication

    doi: 10.1371/journal.pone.0001887

    Figure Lengend Snippet: Treatment with MG132 activates stress. (A) Inhibition of proteasome activated hsp70 synthesis. Control HeLa cells, cells treated for 4 h with MG132, and 4 h VSV-infected cells were incubated for last 30 min with S 35 methionine/cysteine. Cytoplasmic proteins were precipitated with anti-hsp70 and anti-P-VSV Abs. Precipitated proteins were analyzed by electrophoresis and autoradiography. (B) MG132 stimulated eIF2α phosphorylation. HeLa cells were treated with 1 µM of thapsigargin for 1 h and with 5 µM of MG132 for 4 h. 10 µg of protein extracts were analyzed with Abs specific for eIF2α and eIF2α- phosphate (eIF2α-P). Hsp70 is a marker of MG132 activated stress. Keratin 18 (K18) is a loading control.

    Article Snippet: The following antibodies were used: anti-VSV P- and N-protein antibodies obtained by immunization of rabbits, anti-protein 3A mouse monoclonal antibodies were the gift from Dr. K. Kirkegaard, anti-protein 3C rabbit antibodies were a gift from Dr. B. L. Semler, anti-poliovirus capsid proteins antibodies obtained by immunization of rabbits with purified poliovirus, anti-p220 eIF4G mouse antibodies were a gift from Dr. T. Pestova, anti-p65-RelA C-terminus rabbit antibodies (Santa Cruz Biotechnology), anti-IκBα rabbit antibodies (Santa Cruz Biotechnology), anti actin rabbit antibodies (Santa Cruz Biotechnology), anti-GCN2 rabbit antibodies (Santa Cruz Biotechnology), and anti-hsp70 rabbit antibodies (Assay Designs/StressGen).

    Techniques: Inhibition, Infection, Incubation, Electrophoresis, Autoradiography, Marker

    Proteasome inhibitors delay the replication of poliovirus. (A) HeLa cells (triangles) and HeLa cells pre-treated for 2 h with 5 µM proteasome inhibitor MG132 (squares) were infected with poliovirus strain Mahoney (MOI = 5) for 1 h. After replacement of medium, the accumulation of virus in medium was estimated by titration. (B) MG132 inhibits TNF-specific degradation of IκBα. Control HeLa cells and HeLa cells pretreated with 5 µM MG132 for 2 h were incubated with 1 ng/ml of human TNF for 20 min. 10 µg of total protein extracts were analyzed with anti-IκBα Abs. (C) HeLa cells and HeLa cells pre-treated with MG132 were infected with poliovirus (MOI = 5) for 1 h. After medium replacement, protein extracts were collected at different times of infection. The accumulation of poliovirus capsid proteins was tested in Western blotting experiments from 10 µg of protein extracts. (D) The protein extracts described in section B were tested with anti-proteins 3C and 3A Abs. The accumulation of poliovirus proteins 3C, 3A and 3AB were detected in 10 µg of protein extracts. (E) Bortezomib treatment attenuated poliovirus replication. HeLa cells were pretreated with Bortezomib for 2 h, then infected and analyzed as described in panels C and D. K18 was a loading control. Hsp70 is a control of Bortezomib activity.

    Journal: PLoS ONE

    Article Title: Different Effect of Proteasome Inhibition on Vesicular Stomatitis Virus and Poliovirus Replication

    doi: 10.1371/journal.pone.0001887

    Figure Lengend Snippet: Proteasome inhibitors delay the replication of poliovirus. (A) HeLa cells (triangles) and HeLa cells pre-treated for 2 h with 5 µM proteasome inhibitor MG132 (squares) were infected with poliovirus strain Mahoney (MOI = 5) for 1 h. After replacement of medium, the accumulation of virus in medium was estimated by titration. (B) MG132 inhibits TNF-specific degradation of IκBα. Control HeLa cells and HeLa cells pretreated with 5 µM MG132 for 2 h were incubated with 1 ng/ml of human TNF for 20 min. 10 µg of total protein extracts were analyzed with anti-IκBα Abs. (C) HeLa cells and HeLa cells pre-treated with MG132 were infected with poliovirus (MOI = 5) for 1 h. After medium replacement, protein extracts were collected at different times of infection. The accumulation of poliovirus capsid proteins was tested in Western blotting experiments from 10 µg of protein extracts. (D) The protein extracts described in section B were tested with anti-proteins 3C and 3A Abs. The accumulation of poliovirus proteins 3C, 3A and 3AB were detected in 10 µg of protein extracts. (E) Bortezomib treatment attenuated poliovirus replication. HeLa cells were pretreated with Bortezomib for 2 h, then infected and analyzed as described in panels C and D. K18 was a loading control. Hsp70 is a control of Bortezomib activity.

    Article Snippet: The following antibodies were used: anti-VSV P- and N-protein antibodies obtained by immunization of rabbits, anti-protein 3A mouse monoclonal antibodies were the gift from Dr. K. Kirkegaard, anti-protein 3C rabbit antibodies were a gift from Dr. B. L. Semler, anti-poliovirus capsid proteins antibodies obtained by immunization of rabbits with purified poliovirus, anti-p220 eIF4G mouse antibodies were a gift from Dr. T. Pestova, anti-p65-RelA C-terminus rabbit antibodies (Santa Cruz Biotechnology), anti-IκBα rabbit antibodies (Santa Cruz Biotechnology), anti actin rabbit antibodies (Santa Cruz Biotechnology), anti-GCN2 rabbit antibodies (Santa Cruz Biotechnology), and anti-hsp70 rabbit antibodies (Assay Designs/StressGen).

    Techniques: Infection, Titration, Incubation, Western Blot, Activity Assay

    Hsc70 and PKCι distribution depend on the integrity of intermediate filaments in small intestine villi of K8 knockout mice. Small intestines were harvested from K8-null mice (C,D,I,J) or from K18 +/– littermates (A,B,E-H), fixed in 10%

    Journal: Journal of Cell Science

    Article Title: Rescue of atypical protein kinase C in epithelia by the cytoskeleton and Hsp70 family chaperones

    doi: 10.1242/jcs.046979

    Figure Lengend Snippet: Hsc70 and PKCι distribution depend on the integrity of intermediate filaments in small intestine villi of K8 knockout mice. Small intestines were harvested from K8-null mice (C,D,I,J) or from K18 +/– littermates (A,B,E-H), fixed in 10%

    Article Snippet: The antibodies used were as follows: mouse monoclonal anti-PKCι (BD Biosciences); rabbit anti-PKCι (Santa Cruz Biotechnology); rabbit anti-phospho-T555 (pT555) PKCι (Biosource Invitrogen); rabbit anti-phospho turn motif aPKC (Epitomics); PKCζ (atypical) pT410 and PKB (Aktα) pT308 (Cell Signaling); mouse anti-V5 (Invitrogen); mouse anti-K8 (Biomeda); anti-pan-cytokeratin (Sigma); anti-K8 TROMA I (Hybridoma Bank); rabbit anti-Hsc70 (constitutive form of Hsp70; Stressgen Bioreagents); rabbit anti-Hsp70 (inducible form of Hsp70; Stressgen Bioreagents); rabbit anti-Hsp70 (against constitutive and inducible forms of Hsp70; Cell Signaling), rabbit polyclonal anti-caspase 3 (detects full length caspase-3 and its large cleavage fragment; Cell Signaling); mouse anti-actin (C4MP; Biomedicals); rabbit anti-PAR6 (Abcam); rabbit anti-PAR3 (Upstate); mouse anti-pan-14.3.3 (clone CG15; Lab Vision); rabbit anti-Pals1 (Upstate) and mouse anti-ZO1 (Zymed Laboratories).

    Techniques: Knock-Out, Mouse Assay

    Hsp70 and/or Hsc70 are necessary to maintain a pool of pT555 PKCι in steady state in CACO-2 cells. (A) Downregulation of Hsp70 by shRNA leads to the decrease in the amount of pT555 and total PKCι protein in CACO-2 cells. At 7 days after

    Journal: Journal of Cell Science

    Article Title: Rescue of atypical protein kinase C in epithelia by the cytoskeleton and Hsp70 family chaperones

    doi: 10.1242/jcs.046979

    Figure Lengend Snippet: Hsp70 and/or Hsc70 are necessary to maintain a pool of pT555 PKCι in steady state in CACO-2 cells. (A) Downregulation of Hsp70 by shRNA leads to the decrease in the amount of pT555 and total PKCι protein in CACO-2 cells. At 7 days after

    Article Snippet: The antibodies used were as follows: mouse monoclonal anti-PKCι (BD Biosciences); rabbit anti-PKCι (Santa Cruz Biotechnology); rabbit anti-phospho-T555 (pT555) PKCι (Biosource Invitrogen); rabbit anti-phospho turn motif aPKC (Epitomics); PKCζ (atypical) pT410 and PKB (Aktα) pT308 (Cell Signaling); mouse anti-V5 (Invitrogen); mouse anti-K8 (Biomeda); anti-pan-cytokeratin (Sigma); anti-K8 TROMA I (Hybridoma Bank); rabbit anti-Hsc70 (constitutive form of Hsp70; Stressgen Bioreagents); rabbit anti-Hsp70 (inducible form of Hsp70; Stressgen Bioreagents); rabbit anti-Hsp70 (against constitutive and inducible forms of Hsp70; Cell Signaling), rabbit polyclonal anti-caspase 3 (detects full length caspase-3 and its large cleavage fragment; Cell Signaling); mouse anti-actin (C4MP; Biomedicals); rabbit anti-PAR6 (Abcam); rabbit anti-PAR3 (Upstate); mouse anti-pan-14.3.3 (clone CG15; Lab Vision); rabbit anti-Pals1 (Upstate) and mouse anti-ZO1 (Zymed Laboratories).

    Techniques: shRNA

    Hsc70 and PKCι distribution depend on the integrity of intermediate filaments in small intestine crypts of K18R89C mice. Small intestines from K18 +/– , K19 –/– , hK18 R89C knockout/knock-in mice (C,D,I,J) (ko) or control

    Journal: Journal of Cell Science

    Article Title: Rescue of atypical protein kinase C in epithelia by the cytoskeleton and Hsp70 family chaperones

    doi: 10.1242/jcs.046979

    Figure Lengend Snippet: Hsc70 and PKCι distribution depend on the integrity of intermediate filaments in small intestine crypts of K18R89C mice. Small intestines from K18 +/– , K19 –/– , hK18 R89C knockout/knock-in mice (C,D,I,J) (ko) or control

    Article Snippet: The antibodies used were as follows: mouse monoclonal anti-PKCι (BD Biosciences); rabbit anti-PKCι (Santa Cruz Biotechnology); rabbit anti-phospho-T555 (pT555) PKCι (Biosource Invitrogen); rabbit anti-phospho turn motif aPKC (Epitomics); PKCζ (atypical) pT410 and PKB (Aktα) pT308 (Cell Signaling); mouse anti-V5 (Invitrogen); mouse anti-K8 (Biomeda); anti-pan-cytokeratin (Sigma); anti-K8 TROMA I (Hybridoma Bank); rabbit anti-Hsc70 (constitutive form of Hsp70; Stressgen Bioreagents); rabbit anti-Hsp70 (inducible form of Hsp70; Stressgen Bioreagents); rabbit anti-Hsp70 (against constitutive and inducible forms of Hsp70; Cell Signaling), rabbit polyclonal anti-caspase 3 (detects full length caspase-3 and its large cleavage fragment; Cell Signaling); mouse anti-actin (C4MP; Biomedicals); rabbit anti-PAR6 (Abcam); rabbit anti-PAR3 (Upstate); mouse anti-pan-14.3.3 (clone CG15; Lab Vision); rabbit anti-Pals1 (Upstate) and mouse anti-ZO1 (Zymed Laboratories).

    Techniques: Mouse Assay, Knock-Out, Knock-In

    Subcellular distribution of PKCι and Hsc70 modified by overexpression and redistribution of keratins. Frozen sections of duodenum from genetically unmodified FVB/n mice (A-D,I-L) or transgenic K8 overexpressors (K8 oe; E-H,M-P) were processed

    Journal: Journal of Cell Science

    Article Title: Rescue of atypical protein kinase C in epithelia by the cytoskeleton and Hsp70 family chaperones

    doi: 10.1242/jcs.046979

    Figure Lengend Snippet: Subcellular distribution of PKCι and Hsc70 modified by overexpression and redistribution of keratins. Frozen sections of duodenum from genetically unmodified FVB/n mice (A-D,I-L) or transgenic K8 overexpressors (K8 oe; E-H,M-P) were processed

    Article Snippet: The antibodies used were as follows: mouse monoclonal anti-PKCι (BD Biosciences); rabbit anti-PKCι (Santa Cruz Biotechnology); rabbit anti-phospho-T555 (pT555) PKCι (Biosource Invitrogen); rabbit anti-phospho turn motif aPKC (Epitomics); PKCζ (atypical) pT410 and PKB (Aktα) pT308 (Cell Signaling); mouse anti-V5 (Invitrogen); mouse anti-K8 (Biomeda); anti-pan-cytokeratin (Sigma); anti-K8 TROMA I (Hybridoma Bank); rabbit anti-Hsc70 (constitutive form of Hsp70; Stressgen Bioreagents); rabbit anti-Hsp70 (inducible form of Hsp70; Stressgen Bioreagents); rabbit anti-Hsp70 (against constitutive and inducible forms of Hsp70; Cell Signaling), rabbit polyclonal anti-caspase 3 (detects full length caspase-3 and its large cleavage fragment; Cell Signaling); mouse anti-actin (C4MP; Biomedicals); rabbit anti-PAR6 (Abcam); rabbit anti-PAR3 (Upstate); mouse anti-pan-14.3.3 (clone CG15; Lab Vision); rabbit anti-Pals1 (Upstate) and mouse anti-ZO1 (Zymed Laboratories).

    Techniques: Modification, Over Expression, Mouse Assay, Transgenic Assay