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  • 86
    Cell Signaling Technology Inc hspa5 grp78 bip
    High fat and ethanol diets activate PERK signaling in pancreatic acinar cells. ( A ) Western blotting analysis for peIF2α, <t>BiP/GRP78</t> (upper band) or β-actin (as a control) in pancreatic tissue of WT or Mist1 −/− mice fed breeder's chow (BC) or LDC-E or LDC-HF diets for 6 weeks. Quantitative analysis of peIF2α ( B ) or BiP/GRP78 ( C ) accumulation from (A) relative to β-actin accumulation. LDC-E and LDC-HF diets lead to significant increases in peIF2α compared to BC diets in WT mice but not in Mist1 −/− tissue. Conversely, while BiP/GRP78 levels are unchanged in WT mice, they decrease upon exposure to LDC diets in Mist1 −/− mice. Groups were compared using One way ANOVA and a Tukey post-hoc tests: *P
    Hspa5 Grp78 Bip, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stressgen Biotechnologies hspa5 bip
    High fat and ethanol diets activate PERK signaling in pancreatic acinar cells. ( A ) Western blotting analysis for peIF2α, <t>BiP/GRP78</t> (upper band) or β-actin (as a control) in pancreatic tissue of WT or Mist1 −/− mice fed breeder's chow (BC) or LDC-E or LDC-HF diets for 6 weeks. Quantitative analysis of peIF2α ( B ) or BiP/GRP78 ( C ) accumulation from (A) relative to β-actin accumulation. LDC-E and LDC-HF diets lead to significant increases in peIF2α compared to BC diets in WT mice but not in Mist1 −/− tissue. Conversely, while BiP/GRP78 levels are unchanged in WT mice, they decrease upon exposure to LDC diets in Mist1 −/− mice. Groups were compared using One way ANOVA and a Tukey post-hoc tests: *P
    Hspa5 Bip, supplied by Stressgen Biotechnologies, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam grp78 bip
    HKH40A affects transcription of <t>GRP78/BiP.</t> ( a ) qPCR analysis of BiP transcripts in HCT-116 and HT-29 cells untreated or treated with 100 nM HKH40A for 24 h. ( b ) Promoter activity in HCT-116 and HT-29 cells transfected with pGL-BiP-P plasmid and treated with 100 nM HKH40A for 24 h (fold change relative to pGL-4.10 empty vector transfected cells)
    Grp78 Bip, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 206 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals grp78 hspa5 antibody
    <t>Grp78</t> Dominant-Negative Version Rescues Aβ Toxicity (A) Lifespan survival curves of flies expressing Aβ or the Aβ Grp78 dominant-negative version in neurons (+RU) and uninduced controls (-RU). (Aβ +RU and Aβ Grp78 dominant-negative +RU are different. p
    Grp78 Hspa5 Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti hspa5 grp78 bip
    <t>Grp78</t> Dominant-Negative Version Rescues Aβ Toxicity (A) Lifespan survival curves of flies expressing Aβ or the Aβ Grp78 dominant-negative version in neurons (+RU) and uninduced controls (-RU). (Aβ +RU and Aβ Grp78 dominant-negative +RU are different. p
    Anti Hspa5 Grp78 Bip, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Enzo Biochem grp78 hspa5 bip
    <t>Grp78</t> Dominant-Negative Version Rescues Aβ Toxicity (A) Lifespan survival curves of flies expressing Aβ or the Aβ Grp78 dominant-negative version in neurons (+RU) and uninduced controls (-RU). (Aβ +RU and Aβ Grp78 dominant-negative +RU are different. p
    Grp78 Hspa5 Bip, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems grp78 hspa5
    <t>Grp78</t> Dominant-Negative Version Rescues Aβ Toxicity (A) Lifespan survival curves of flies expressing Aβ or the Aβ Grp78 dominant-negative version in neurons (+RU) and uninduced controls (-RU). (Aβ +RU and Aβ Grp78 dominant-negative +RU are different. p
    Grp78 Hspa5, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology grp78 bip
    ERp72 and <t>GRP78/BiP</t> protein are elevated and ERp72 oxidized state is increased in AdDNPerk 832/13 beta cells . A. Triplicate samples were probed with antibodies to ERp72, ERp57, BiP and actin, which served as the loading control. B. ERp72 expression was increased in islets from P2 Perk KO mice. Samples are from duplicate mice for each genotype. C. At 24 hr or 36 hr post-transduction, protein samples were isolated and treated with AMS to differentiate the reduced and oxidized forms of ERp72 and ERp57 on PAGE gels. Western blots showed increased oxidized isoforms of both ERp72 and ERp57 in 832/13 cells infected with AdDNPerk compared to cells infected with AdLacZ .
    Grp78 Bip, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson bip grp78
    Deletion of Tlr2 inhibits increased cytokine sensitivity in Trim58 -deficient myeloid cells. ( A ) Assessment of TLR2, mature IL-1β, arginase-1, and <t>BIP/GRP78</t> protein synthesis in peritoneal myeloid (PM) cells from male Trim58 +/+ and Trim58 −/− mice ( n = 4–6 per genotype) stimulated with TNF-α (100 ng/ml) and IFN-γ (5 ng/ml) for 24.5 h by immunoblot (IB) analysis. ( B ) Assessment of mature IL-1β protein synthesis in PM cells from male Tlr2 −/− , Trim58 −/− and Trim58 −/− Tlr2 −/− mice ( n = 4–6/genotype) stimulated with TNF-α (100 ng/ml) and IFN-γ (5 ng/ml) for 24.5 h by IB analysis. Blots were reprobed with anti-GAPDH to confirm equal loading. Right margin, molecular size markers (kDa). Representative results of at least two independent experiments are shown.
    Bip Grp78, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 101 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology bip grp78
    Microscopic analysis of the distribution of <t>BiP/GRP78</t> and CHOP/Gadd153 in immunohistochemically stained cochleae (SP, ×400). (a-1–a-3) Cells that were positively stained for BiP/GRP were located in the organ of Corti, the lateral wall, and the spiral ganglion, respectively, in the 1d group (positive expression: immuoreactivity is brown versus the blue hematoxylin stain, indicated by ↑). (b-1–b-3) Cells that were positively stained for CHOP/Gadd153 were located in the same regions as those shown in (a), in the 1d group (positive expression: immunoreactivity is tan or medium-brown particles versus hematoxylin-stain, indicated by ↑). (c-1–c-3) Negative control incubated with PBS instead of the primary antibodies (organ of Corti, the lateral wall, and the spiral ganglion, respectively)
    Bip Grp78, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson grp78 bip
    <t>Grp78/BiP</t> protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p
    Grp78 Bip, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti hspa5 bip
    <t>Grp78/BiP</t> protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p
    Anti Hspa5 Bip, supplied by Cell Signaling Technology Inc, 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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    Abcam hspa5 grp78
    <t>Grp78/BiP</t> protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p
    Hspa5 Grp78, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam recombinant bip protein
    <t>Grp78/BiP</t> protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p
    Recombinant Bip Protein, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech anti hspa5 grp78
    <t>Grp78/BiP</t> protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p
    Anti Hspa5 Grp78, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    StressMarq anti hspa5 grp78
    <t>Grp78/BiP</t> protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p
    Anti Hspa5 Grp78, supplied by StressMarq, 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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    Thermo Fisher bip grp78 antiserum
    <t>Grp78/BiP</t> protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p
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    Image Search Results


    High fat and ethanol diets activate PERK signaling in pancreatic acinar cells. ( A ) Western blotting analysis for peIF2α, BiP/GRP78 (upper band) or β-actin (as a control) in pancreatic tissue of WT or Mist1 −/− mice fed breeder's chow (BC) or LDC-E or LDC-HF diets for 6 weeks. Quantitative analysis of peIF2α ( B ) or BiP/GRP78 ( C ) accumulation from (A) relative to β-actin accumulation. LDC-E and LDC-HF diets lead to significant increases in peIF2α compared to BC diets in WT mice but not in Mist1 −/− tissue. Conversely, while BiP/GRP78 levels are unchanged in WT mice, they decrease upon exposure to LDC diets in Mist1 −/− mice. Groups were compared using One way ANOVA and a Tukey post-hoc tests: *P

    Journal: PLoS ONE

    Article Title: The Absence of MIST1 Leads to Increased Ethanol Sensitivity and Decreased Activity of the Unfolded Protein Response in Mouse Pancreatic Acinar Cells

    doi: 10.1371/journal.pone.0028863

    Figure Lengend Snippet: High fat and ethanol diets activate PERK signaling in pancreatic acinar cells. ( A ) Western blotting analysis for peIF2α, BiP/GRP78 (upper band) or β-actin (as a control) in pancreatic tissue of WT or Mist1 −/− mice fed breeder's chow (BC) or LDC-E or LDC-HF diets for 6 weeks. Quantitative analysis of peIF2α ( B ) or BiP/GRP78 ( C ) accumulation from (A) relative to β-actin accumulation. LDC-E and LDC-HF diets lead to significant increases in peIF2α compared to BC diets in WT mice but not in Mist1 −/− tissue. Conversely, while BiP/GRP78 levels are unchanged in WT mice, they decrease upon exposure to LDC diets in Mist1 −/− mice. Groups were compared using One way ANOVA and a Tukey post-hoc tests: *P

    Article Snippet: Another target of ATF6α, BiP/GRP78 also decreased in LDC-E and LDC-HF fed Mist1−/− mice , supporting the findings that Mist1−/− mice had a reduction in ATF6 signaling as a result of ethanol or high-fat consumption.

    Techniques: Western Blot, Mouse Assay

    Mist1 −/− pancreatic tissue exhibits increased activation of the UPR. ( A ) Western blot analysis of key UPR markers in 2 month old WT and Mist1 −/− whole pancreatic lysates. Mist1 −/− extracts show significantly increased accumulations of BiP/GRP78 ( B ), GADD34 ( C ) and sXBP1 ( D ), but not peIF2α (p = 0.743) or uXBP1 (p = 0.532) relative to WT pancreatic tissue. *P

    Journal: PLoS ONE

    Article Title: The Absence of MIST1 Leads to Increased Ethanol Sensitivity and Decreased Activity of the Unfolded Protein Response in Mouse Pancreatic Acinar Cells

    doi: 10.1371/journal.pone.0028863

    Figure Lengend Snippet: Mist1 −/− pancreatic tissue exhibits increased activation of the UPR. ( A ) Western blot analysis of key UPR markers in 2 month old WT and Mist1 −/− whole pancreatic lysates. Mist1 −/− extracts show significantly increased accumulations of BiP/GRP78 ( B ), GADD34 ( C ) and sXBP1 ( D ), but not peIF2α (p = 0.743) or uXBP1 (p = 0.532) relative to WT pancreatic tissue. *P

    Article Snippet: Another target of ATF6α, BiP/GRP78 also decreased in LDC-E and LDC-HF fed Mist1−/− mice , supporting the findings that Mist1−/− mice had a reduction in ATF6 signaling as a result of ethanol or high-fat consumption.

    Techniques: Activation Assay, Western Blot

    Positions of Grp78/BiP positive staining cells in lamellar tissue. All images are from laminitic samples. Grp78/BiP (green) and WGA (red) were localized as described in Methods. Images were selected to show the range of Grp78/BiP staining patterns observed in laminitic tissues. In the Abaxial region, Grp78/BiP expressing cells (yellow arrows) are found in multiple layers of suprabasal cells, or limited to a single layer of cells adjacent to the keratinized axis. In the middle region, Grp78/BiP positive cells (yellow arrows) are found adjacent to the keratinized axis (KA), often several cell layers deep. In some cases, positive cells are found along the length of the keratinized axis, while in other tissues expression is limited to groups of cells. All images are shown at the same magnification. Scale bar is 50 μm

    Journal: BMC Veterinary Research

    Article Title: Detection of endoplasmic reticulum stress and the unfolded protein response in naturally-occurring endocrinopathic equine laminitis

    doi: 10.1186/s12917-018-1748-x

    Figure Lengend Snippet: Positions of Grp78/BiP positive staining cells in lamellar tissue. All images are from laminitic samples. Grp78/BiP (green) and WGA (red) were localized as described in Methods. Images were selected to show the range of Grp78/BiP staining patterns observed in laminitic tissues. In the Abaxial region, Grp78/BiP expressing cells (yellow arrows) are found in multiple layers of suprabasal cells, or limited to a single layer of cells adjacent to the keratinized axis. In the middle region, Grp78/BiP positive cells (yellow arrows) are found adjacent to the keratinized axis (KA), often several cell layers deep. In some cases, positive cells are found along the length of the keratinized axis, while in other tissues expression is limited to groups of cells. All images are shown at the same magnification. Scale bar is 50 μm

    Article Snippet: Paraffin-embedded 6 μm thick sections of formalin-fixed tissues were used for immunofluorescent localization of Grp78/BiP (Cell Signaling Technology clone C50B12).

    Techniques: Staining, Whole Genome Amplification, Expressing

    Grp78/BiP is expressed at higher levels in laminitic tissues. a . Representative Grp78/BiP immunoblot, reprobed for alpha-tubulin as a loading control. A band of ~ 78 kd co-migrates with the Hela (human) protein. b : Legend to Figure S1A)). Lanes in (A,B) are labeled as: control (C), EL front limb (F) and EL hind limb (H). ( c ). Box plot of normalized Grp78/BiP band intensities in control ( n = 6), EL front limbs (=12) and EL hind limbs ( n = 8). Horizontal lines represent the median value for each group. Grp78/BiP expression was not significantly different between controls and EL hind limbs ( p = 0.77) d . Comparison of Grp78/BiP expression in paired samples ( n = 8) from EL front and hind limbs from the same horses. The inset shows the 4 sample pairs with small differences in Grp78/BiP band intensities between EL front and hind limb samples. The same pattern is apparent in 3 of 4 pairs, where the value for Grp78/BiP level in the front limb is greater than that in the hind limb

    Journal: BMC Veterinary Research

    Article Title: Detection of endoplasmic reticulum stress and the unfolded protein response in naturally-occurring endocrinopathic equine laminitis

    doi: 10.1186/s12917-018-1748-x

    Figure Lengend Snippet: Grp78/BiP is expressed at higher levels in laminitic tissues. a . Representative Grp78/BiP immunoblot, reprobed for alpha-tubulin as a loading control. A band of ~ 78 kd co-migrates with the Hela (human) protein. b : Legend to Figure S1A)). Lanes in (A,B) are labeled as: control (C), EL front limb (F) and EL hind limb (H). ( c ). Box plot of normalized Grp78/BiP band intensities in control ( n = 6), EL front limbs (=12) and EL hind limbs ( n = 8). Horizontal lines represent the median value for each group. Grp78/BiP expression was not significantly different between controls and EL hind limbs ( p = 0.77) d . Comparison of Grp78/BiP expression in paired samples ( n = 8) from EL front and hind limbs from the same horses. The inset shows the 4 sample pairs with small differences in Grp78/BiP band intensities between EL front and hind limb samples. The same pattern is apparent in 3 of 4 pairs, where the value for Grp78/BiP level in the front limb is greater than that in the hind limb

    Article Snippet: Paraffin-embedded 6 μm thick sections of formalin-fixed tissues were used for immunofluorescent localization of Grp78/BiP (Cell Signaling Technology clone C50B12).

    Techniques: Labeling, Expressing

    Localization of Grp78/BiP to suprabasal keratinocytes of the epidermal lamellae in laminitic tissue. Tissue sections were stained as described in Methods to localize Grp78/BiP (green) and wheat germ agglutinin (WGA; red) to outline cell plasma membranes (epidermis) and extracellular matrix (dermis). The Grp78/BiP channel is also shown separately in grayscale for each image. Representative images from Abaxial (AbAx), Middle (Mid) and Axial positions (Ax) are shown for control and laminitic samples. The control tissues showed no significant Grp78/BiP staining under the conditions used. In contrast, laminitic tissues showed bright Grp78/BiP expression in suprabasal epidermal keratinocytes located at the abaxial region and along the keratinized axis. Positive staining was absent from SELs in the axial region. All images are shown at the same magnification. Scale bar is 50 μm

    Journal: BMC Veterinary Research

    Article Title: Detection of endoplasmic reticulum stress and the unfolded protein response in naturally-occurring endocrinopathic equine laminitis

    doi: 10.1186/s12917-018-1748-x

    Figure Lengend Snippet: Localization of Grp78/BiP to suprabasal keratinocytes of the epidermal lamellae in laminitic tissue. Tissue sections were stained as described in Methods to localize Grp78/BiP (green) and wheat germ agglutinin (WGA; red) to outline cell plasma membranes (epidermis) and extracellular matrix (dermis). The Grp78/BiP channel is also shown separately in grayscale for each image. Representative images from Abaxial (AbAx), Middle (Mid) and Axial positions (Ax) are shown for control and laminitic samples. The control tissues showed no significant Grp78/BiP staining under the conditions used. In contrast, laminitic tissues showed bright Grp78/BiP expression in suprabasal epidermal keratinocytes located at the abaxial region and along the keratinized axis. Positive staining was absent from SELs in the axial region. All images are shown at the same magnification. Scale bar is 50 μm

    Article Snippet: Paraffin-embedded 6 μm thick sections of formalin-fixed tissues were used for immunofluorescent localization of Grp78/BiP (Cell Signaling Technology clone C50B12).

    Techniques: Staining, Whole Genome Amplification, Expressing

    Dexamethasone treatment reduces incidence of pseudoimplantation and NF κ B-mediated inflammation in Msx1/Msx2 d/d uteri under delayed conditions. A , incidence of pseudoimplantation sites ( pseudoIS ) in vehicle- ( Veh ), celecoxib- ( Cele ), or dexamethasone-treated ( Dex ) Msx1/Msx2 d/d uteri under delayed conditions. Bars showing color-coded segments represent the percentage of mice with pseudoIS ( red ), blue bands ( green ), and recovered blastocysts ( blue ). Although percentages of pseudoIS with faint or no blue bands at the site of blastocysts are shown within parentheses in the red segments of bars , numbers of mice used in each set of experiments are shown above the bars. B , representative images of Msx1/Msx2 d/d uteri after vehicle or Dex treatment as compared with dexamethasone-treated floxed uteri. Arrow , faint blue band. C , left panel , Western blotting results showing reduced levels of pIκB, α-subunits of the 20S proteasome, p-eIF2α, and HSPA5/Bip in Msx1/Msx2 d/d mice after dexamethasone treatment. Right panels , quantification of Western blotting results (means ± S.E.). *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Muscle Segment Homeobox Genes Direct Embryonic Diapause by Limiting Inflammation in the Uterus *

    doi: 10.1074/jbc.M115.655001

    Figure Lengend Snippet: Dexamethasone treatment reduces incidence of pseudoimplantation and NF κ B-mediated inflammation in Msx1/Msx2 d/d uteri under delayed conditions. A , incidence of pseudoimplantation sites ( pseudoIS ) in vehicle- ( Veh ), celecoxib- ( Cele ), or dexamethasone-treated ( Dex ) Msx1/Msx2 d/d uteri under delayed conditions. Bars showing color-coded segments represent the percentage of mice with pseudoIS ( red ), blue bands ( green ), and recovered blastocysts ( blue ). Although percentages of pseudoIS with faint or no blue bands at the site of blastocysts are shown within parentheses in the red segments of bars , numbers of mice used in each set of experiments are shown above the bars. B , representative images of Msx1/Msx2 d/d uteri after vehicle or Dex treatment as compared with dexamethasone-treated floxed uteri. Arrow , faint blue band. C , left panel , Western blotting results showing reduced levels of pIκB, α-subunits of the 20S proteasome, p-eIF2α, and HSPA5/Bip in Msx1/Msx2 d/d mice after dexamethasone treatment. Right panels , quantification of Western blotting results (means ± S.E.). *, p

    Article Snippet: Antibodies to phosphorylated IκB (mouse, Cell Signaling Technology, 9246), total IκB (rabbit, Cell Signaling Technology, 9242), 20S proteasome α-1, 2, 3, 5, 6, 7 subunits (mouse, Enzo, MCP231), α6 (rabbit, laboratory-generated), Rpt6 (mouse, laboratory-generated), Rpn8 (rabbit, laboratory-generated), HSPA5/Bip (goat, sc-1050), phosphorylated eIF2α (rabbit, Cell Signaling, 3597S), total eIF2α (rabbit, sc-11386), ubiquitin lysine 48 (rabbit, Millipore, 05-1307), and ubiquitin lysine 63 (rabbit, Millipore, 05-1308) were used as previously described ( ).

    Techniques: Mouse Assay, Western Blot

    ER stress is evident in Msx1/Msx2 d/d uteri under delayed conditions and is aggravated by bortezomib treatment. A , representative results of immunofluorescence showing apical epithelial expression of HSPA5/Bip in Msx1/Msx2 f/f uteri, with increased intensity and dispersed Bip-positive stromal cells in Msx1/Msx2 d/d uteri. Stromal Bip expression is widespread at the pseudoimplantation site ( Pseudo-IS ). Arrowheads , dispersed Bip-positive cells. Bar , 250 μm. B , blastocysts recovery ( top bar graph ) from Msx1/Msx2 d/d after bortezomib treatment females was significantly lower with enhanced rate of pseudoimplantation sites ( bottom bar graph ) compared with floxed littermates on day 10 (means ± S.E.). Numbers in parentheses indicate number of females that produced blastocysts or pseudoimplantation sites, respectively, compared with the number of females assessed.

    Journal: The Journal of Biological Chemistry

    Article Title: Muscle Segment Homeobox Genes Direct Embryonic Diapause by Limiting Inflammation in the Uterus *

    doi: 10.1074/jbc.M115.655001

    Figure Lengend Snippet: ER stress is evident in Msx1/Msx2 d/d uteri under delayed conditions and is aggravated by bortezomib treatment. A , representative results of immunofluorescence showing apical epithelial expression of HSPA5/Bip in Msx1/Msx2 f/f uteri, with increased intensity and dispersed Bip-positive stromal cells in Msx1/Msx2 d/d uteri. Stromal Bip expression is widespread at the pseudoimplantation site ( Pseudo-IS ). Arrowheads , dispersed Bip-positive cells. Bar , 250 μm. B , blastocysts recovery ( top bar graph ) from Msx1/Msx2 d/d after bortezomib treatment females was significantly lower with enhanced rate of pseudoimplantation sites ( bottom bar graph ) compared with floxed littermates on day 10 (means ± S.E.). Numbers in parentheses indicate number of females that produced blastocysts or pseudoimplantation sites, respectively, compared with the number of females assessed.

    Article Snippet: Antibodies to phosphorylated IκB (mouse, Cell Signaling Technology, 9246), total IκB (rabbit, Cell Signaling Technology, 9242), 20S proteasome α-1, 2, 3, 5, 6, 7 subunits (mouse, Enzo, MCP231), α6 (rabbit, laboratory-generated), Rpt6 (mouse, laboratory-generated), Rpn8 (rabbit, laboratory-generated), HSPA5/Bip (goat, sc-1050), phosphorylated eIF2α (rabbit, Cell Signaling, 3597S), total eIF2α (rabbit, sc-11386), ubiquitin lysine 48 (rabbit, Millipore, 05-1307), and ubiquitin lysine 63 (rabbit, Millipore, 05-1308) were used as previously described ( ).

    Techniques: Immunofluorescence, Expressing, Produced

    HKH40A affects transcription of GRP78/BiP. ( a ) qPCR analysis of BiP transcripts in HCT-116 and HT-29 cells untreated or treated with 100 nM HKH40A for 24 h. ( b ) Promoter activity in HCT-116 and HT-29 cells transfected with pGL-BiP-P plasmid and treated with 100 nM HKH40A for 24 h (fold change relative to pGL-4.10 empty vector transfected cells)

    Journal: Cell Death & Disease

    Article Title: HKH40A downregulates GRP78/BiP expression in cancer cells

    doi: 10.1038/cddis.2014.203

    Figure Lengend Snippet: HKH40A affects transcription of GRP78/BiP. ( a ) qPCR analysis of BiP transcripts in HCT-116 and HT-29 cells untreated or treated with 100 nM HKH40A for 24 h. ( b ) Promoter activity in HCT-116 and HT-29 cells transfected with pGL-BiP-P plasmid and treated with 100 nM HKH40A for 24 h (fold change relative to pGL-4.10 empty vector transfected cells)

    Article Snippet: After 1 h blocking of nonspecific binding sites using 5% bovine serum albumin (BSA), cells were stained 1 h with the primary antibody to GRP78/BiP (Rabbit polyclonal, ab21685, Abcam) diluted 1 : 500 in 5% BSA then washed with PBS-T, stained with secondary antibody Alexa 594 (1 : 1000 in 5% BSA) for 1 h followed by final PBS-T washes and examined using a Zeiss LC510 confocal microscope.

    Techniques: Real-time Polymerase Chain Reaction, Activity Assay, Transfection, Plasmid Preparation

    HKH40A directly binds BiP and affects its stability. ( a ) Colocalization of HKH40A with BiP. HCT-116 cells were treated with 100 nM HKH40A for 3 h, fixed and immunostained for GRP78. The drug with intrinistic ability to fluoresce binds to DNA and emits very bright fluorescence. Fluorescence is also visible in cytoplasm. Immunostaining of cells with GRP78 shows perinuclear and cytoplasmic localization. Some overlap of the two signals in the perinuclear region can be noted in the orthogonal views of the Z-stacks, scale bar 10 μ m. ( b ) Direct interaction of HKH40A with recombinant human GRP78/BiP. Microscale thermophoresis analysis revealed direct interaction of HKH40A with recombinant human GRP78/BiP. Analysis was performed using the intrinsic ability of HKH40A to fluoresce. Purified recombinant human K-Ras (1–166) was used as a negative control (in red). Its addition caused no change in the thermophoretic mobility of the compound. ( c ) Proteosomal degradation is involved in HKH40A downregulation of BiP. HCT-116 and HT-29 cells were treated with 100 nM HKH40A, 10 μ M MG132 or a combination of both agents for 6 h, then drugs were removed, cells were cultured in a drug-free medium for up to 24 h, collected and analyzed by western blot. ( d ) Topoisomerase1 and topoisomerase2 inhibition is not responsible for the reduction of GRP78/BiP level by HKH40A. HT-29 cells were cultured for 24 h with vehicle (1), 100 nM HKH40A (2), 5 μ M CPT-11 (3), 10 μ M Etoposide (4), 250 nM adriamycin (5) and 1 μ M WMC26 (6). Total cell lysates were subjected to western blots with anti-GRP78/BiP. β -Actin was used as a loading control

    Journal: Cell Death & Disease

    Article Title: HKH40A downregulates GRP78/BiP expression in cancer cells

    doi: 10.1038/cddis.2014.203

    Figure Lengend Snippet: HKH40A directly binds BiP and affects its stability. ( a ) Colocalization of HKH40A with BiP. HCT-116 cells were treated with 100 nM HKH40A for 3 h, fixed and immunostained for GRP78. The drug with intrinistic ability to fluoresce binds to DNA and emits very bright fluorescence. Fluorescence is also visible in cytoplasm. Immunostaining of cells with GRP78 shows perinuclear and cytoplasmic localization. Some overlap of the two signals in the perinuclear region can be noted in the orthogonal views of the Z-stacks, scale bar 10 μ m. ( b ) Direct interaction of HKH40A with recombinant human GRP78/BiP. Microscale thermophoresis analysis revealed direct interaction of HKH40A with recombinant human GRP78/BiP. Analysis was performed using the intrinsic ability of HKH40A to fluoresce. Purified recombinant human K-Ras (1–166) was used as a negative control (in red). Its addition caused no change in the thermophoretic mobility of the compound. ( c ) Proteosomal degradation is involved in HKH40A downregulation of BiP. HCT-116 and HT-29 cells were treated with 100 nM HKH40A, 10 μ M MG132 or a combination of both agents for 6 h, then drugs were removed, cells were cultured in a drug-free medium for up to 24 h, collected and analyzed by western blot. ( d ) Topoisomerase1 and topoisomerase2 inhibition is not responsible for the reduction of GRP78/BiP level by HKH40A. HT-29 cells were cultured for 24 h with vehicle (1), 100 nM HKH40A (2), 5 μ M CPT-11 (3), 10 μ M Etoposide (4), 250 nM adriamycin (5) and 1 μ M WMC26 (6). Total cell lysates were subjected to western blots with anti-GRP78/BiP. β -Actin was used as a loading control

    Article Snippet: After 1 h blocking of nonspecific binding sites using 5% bovine serum albumin (BSA), cells were stained 1 h with the primary antibody to GRP78/BiP (Rabbit polyclonal, ab21685, Abcam) diluted 1 : 500 in 5% BSA then washed with PBS-T, stained with secondary antibody Alexa 594 (1 : 1000 in 5% BSA) for 1 h followed by final PBS-T washes and examined using a Zeiss LC510 confocal microscope.

    Techniques: Fluorescence, Immunostaining, Recombinant, Microscale Thermophoresis, Purification, Negative Control, Cell Culture, Western Blot, Inhibition, Cycling Probe Technology

    Grp78 Dominant-Negative Version Rescues Aβ Toxicity (A) Lifespan survival curves of flies expressing Aβ or the Aβ Grp78 dominant-negative version in neurons (+RU) and uninduced controls (-RU). (Aβ +RU and Aβ Grp78 dominant-negative +RU are different. p

    Journal: Current Biology

    Article Title: Increased Glucose Transport into Neurons Rescues Aβ Toxicity in Drosophila

    doi: 10.1016/j.cub.2016.07.017

    Figure Lengend Snippet: Grp78 Dominant-Negative Version Rescues Aβ Toxicity (A) Lifespan survival curves of flies expressing Aβ or the Aβ Grp78 dominant-negative version in neurons (+RU) and uninduced controls (-RU). (Aβ +RU and Aβ Grp78 dominant-negative +RU are different. p

    Article Snippet: Primary antibody dilutions used were as follows: anti-Grp78, 1:1,000 (Novus Biologicals, NBP1-06274); anti-actin, 1:10,000 (Abcam, ab1801); anti-Ubiquitin, 1:1,000 (Millipore, FK2); and anti-eIF2A-phospho, 1:1,000 (Cell Signaling, 3597).

    Techniques: Dominant Negative Mutation, Expressing

    UPR Components Activated in Aβ-Expressing Flies Are Induced Even Further by Glut1 Overexpression (A) Western blot of eIF2 phosphorylation levels in heads of Aβ- and AβGlut1-expressing flies (+RU) and in controls (-RU), showing no significant difference. Bottom: plotted as means ± SEM (n = 3). Top: a representative gel from the same samples. (B) Grp78 mRNA levels in heads of 18-day-old flies expressing Aβ or Aβ Glut1 in neurons (+RU) and uninduced controls (-RU), measured by qPCR (relative to eIF1A), plotted as means ± SEM. Genotypes: UAS Aβ; elavGS , UAS Aβ/UAS Glut1; elavGS . (C) Quantification of GFP fluorescence in fly brains expressing an Xbp1GFP splicing reporter, plotted as means ± SEM (n = 6–13). Genotypes: elavGS/UAS-Xbp1GFP , UAS Aβ; elavGS/UAS-Xbp1GFP , UAS Aβ/UAS Glut1; elavGS/UAS-Xbp1GFP . (D) Spliced Xbp1 mRNA levels in heads of 18-day-old flies expressing Aβ or Aβ Glut1 in neurons (+RU) and uninduced controls (-RU), measured by qPCR (relative to eIF1A), plotted as means ± SEM (E) Western blot of Grp78 in 14-day-old flies of the same genotypes, plotted below as means ± SEM (n = 6–16). The image is a representative gel of the same samples. Genotypes: UAS Aβ; elavGS , UAS Aβ/UAS Glut1; elavGS . ∗ p ≤ 0.05; ∗∗ p ≤ 0.01, by ANOVA. See also Figure S2 .

    Journal: Current Biology

    Article Title: Increased Glucose Transport into Neurons Rescues Aβ Toxicity in Drosophila

    doi: 10.1016/j.cub.2016.07.017

    Figure Lengend Snippet: UPR Components Activated in Aβ-Expressing Flies Are Induced Even Further by Glut1 Overexpression (A) Western blot of eIF2 phosphorylation levels in heads of Aβ- and AβGlut1-expressing flies (+RU) and in controls (-RU), showing no significant difference. Bottom: plotted as means ± SEM (n = 3). Top: a representative gel from the same samples. (B) Grp78 mRNA levels in heads of 18-day-old flies expressing Aβ or Aβ Glut1 in neurons (+RU) and uninduced controls (-RU), measured by qPCR (relative to eIF1A), plotted as means ± SEM. Genotypes: UAS Aβ; elavGS , UAS Aβ/UAS Glut1; elavGS . (C) Quantification of GFP fluorescence in fly brains expressing an Xbp1GFP splicing reporter, plotted as means ± SEM (n = 6–13). Genotypes: elavGS/UAS-Xbp1GFP , UAS Aβ; elavGS/UAS-Xbp1GFP , UAS Aβ/UAS Glut1; elavGS/UAS-Xbp1GFP . (D) Spliced Xbp1 mRNA levels in heads of 18-day-old flies expressing Aβ or Aβ Glut1 in neurons (+RU) and uninduced controls (-RU), measured by qPCR (relative to eIF1A), plotted as means ± SEM (E) Western blot of Grp78 in 14-day-old flies of the same genotypes, plotted below as means ± SEM (n = 6–16). The image is a representative gel of the same samples. Genotypes: UAS Aβ; elavGS , UAS Aβ/UAS Glut1; elavGS . ∗ p ≤ 0.05; ∗∗ p ≤ 0.01, by ANOVA. See also Figure S2 .

    Article Snippet: Primary antibody dilutions used were as follows: anti-Grp78, 1:1,000 (Novus Biologicals, NBP1-06274); anti-actin, 1:10,000 (Abcam, ab1801); anti-Ubiquitin, 1:1,000 (Millipore, FK2); and anti-eIF2A-phospho, 1:1,000 (Cell Signaling, 3597).

    Techniques: Expressing, Over Expression, Western Blot, Real-time Polymerase Chain Reaction, Fluorescence

    ERp72 and GRP78/BiP protein are elevated and ERp72 oxidized state is increased in AdDNPerk 832/13 beta cells . A. Triplicate samples were probed with antibodies to ERp72, ERp57, BiP and actin, which served as the loading control. B. ERp72 expression was increased in islets from P2 Perk KO mice. Samples are from duplicate mice for each genotype. C. At 24 hr or 36 hr post-transduction, protein samples were isolated and treated with AMS to differentiate the reduced and oxidized forms of ERp72 and ERp57 on PAGE gels. Western blots showed increased oxidized isoforms of both ERp72 and ERp57 in 832/13 cells infected with AdDNPerk compared to cells infected with AdLacZ .

    Journal: BMC Cell Biology

    Article Title: Acute ablation of PERK results in ER dysfunctions followed by reduced insulin secretion and cell proliferation

    doi: 10.1186/1471-2121-10-61

    Figure Lengend Snippet: ERp72 and GRP78/BiP protein are elevated and ERp72 oxidized state is increased in AdDNPerk 832/13 beta cells . A. Triplicate samples were probed with antibodies to ERp72, ERp57, BiP and actin, which served as the loading control. B. ERp72 expression was increased in islets from P2 Perk KO mice. Samples are from duplicate mice for each genotype. C. At 24 hr or 36 hr post-transduction, protein samples were isolated and treated with AMS to differentiate the reduced and oxidized forms of ERp72 and ERp57 on PAGE gels. Western blots showed increased oxidized isoforms of both ERp72 and ERp57 in 832/13 cells infected with AdDNPerk compared to cells infected with AdLacZ .

    Article Snippet: Primary antibodies used in the analysis were: ERp72 (1:2500, Stressgen, Inc), GRP78/BiP (1:500, Santa Cruz, Inc), ERp57 (1:300, Santa Cruz), and anti-GFP (Sigma, Inc.) to detect EFYP-ATF6.

    Techniques: Expressing, Mouse Assay, Transduction, Isolation, Affinity Magnetic Separation, Polyacrylamide Gel Electrophoresis, Western Blot, Infection

    Deletion of Tlr2 inhibits increased cytokine sensitivity in Trim58 -deficient myeloid cells. ( A ) Assessment of TLR2, mature IL-1β, arginase-1, and BIP/GRP78 protein synthesis in peritoneal myeloid (PM) cells from male Trim58 +/+ and Trim58 −/− mice ( n = 4–6 per genotype) stimulated with TNF-α (100 ng/ml) and IFN-γ (5 ng/ml) for 24.5 h by immunoblot (IB) analysis. ( B ) Assessment of mature IL-1β protein synthesis in PM cells from male Tlr2 −/− , Trim58 −/− and Trim58 −/− Tlr2 −/− mice ( n = 4–6/genotype) stimulated with TNF-α (100 ng/ml) and IFN-γ (5 ng/ml) for 24.5 h by IB analysis. Blots were reprobed with anti-GAPDH to confirm equal loading. Right margin, molecular size markers (kDa). Representative results of at least two independent experiments are shown.

    Journal: The Journal of Immunology Author Choice

    Article Title: TRIM58 Restrains Intestinal Mucosal Inflammation by Negatively Regulating TLR2 in Myeloid Cells

    doi: 10.4049/jimmunol.1900413

    Figure Lengend Snippet: Deletion of Tlr2 inhibits increased cytokine sensitivity in Trim58 -deficient myeloid cells. ( A ) Assessment of TLR2, mature IL-1β, arginase-1, and BIP/GRP78 protein synthesis in peritoneal myeloid (PM) cells from male Trim58 +/+ and Trim58 −/− mice ( n = 4–6 per genotype) stimulated with TNF-α (100 ng/ml) and IFN-γ (5 ng/ml) for 24.5 h by immunoblot (IB) analysis. ( B ) Assessment of mature IL-1β protein synthesis in PM cells from male Tlr2 −/− , Trim58 −/− and Trim58 −/− Tlr2 −/− mice ( n = 4–6/genotype) stimulated with TNF-α (100 ng/ml) and IFN-γ (5 ng/ml) for 24.5 h by IB analysis. Blots were reprobed with anti-GAPDH to confirm equal loading. Right margin, molecular size markers (kDa). Representative results of at least two independent experiments are shown.

    Article Snippet: CD3e (SP7) Ab was from Thermo Fisher Scientific, BIP/GRP78 (40/BiP) from BD Biosciences, IL-1β from Abcam (catalog no. ab9722), and HA (H6908) was from Merck.

    Techniques: Mouse Assay

    Microscopic analysis of the distribution of BiP/GRP78 and CHOP/Gadd153 in immunohistochemically stained cochleae (SP, ×400). (a-1–a-3) Cells that were positively stained for BiP/GRP were located in the organ of Corti, the lateral wall, and the spiral ganglion, respectively, in the 1d group (positive expression: immuoreactivity is brown versus the blue hematoxylin stain, indicated by ↑). (b-1–b-3) Cells that were positively stained for CHOP/Gadd153 were located in the same regions as those shown in (a), in the 1d group (positive expression: immunoreactivity is tan or medium-brown particles versus hematoxylin-stain, indicated by ↑). (c-1–c-3) Negative control incubated with PBS instead of the primary antibodies (organ of Corti, the lateral wall, and the spiral ganglion, respectively)

    Journal: Noise & Health

    Article Title: The Protective Effect of the Endoplasmic Reticulum Stress-Related Factors BiP/GRP78 and CHOP/Gadd153 on Noise-induced Hearing Loss in Guinea Pigs

    doi: 10.4103/1463-1741.192481

    Figure Lengend Snippet: Microscopic analysis of the distribution of BiP/GRP78 and CHOP/Gadd153 in immunohistochemically stained cochleae (SP, ×400). (a-1–a-3) Cells that were positively stained for BiP/GRP were located in the organ of Corti, the lateral wall, and the spiral ganglion, respectively, in the 1d group (positive expression: immuoreactivity is brown versus the blue hematoxylin stain, indicated by ↑). (b-1–b-3) Cells that were positively stained for CHOP/Gadd153 were located in the same regions as those shown in (a), in the 1d group (positive expression: immunoreactivity is tan or medium-brown particles versus hematoxylin-stain, indicated by ↑). (c-1–c-3) Negative control incubated with PBS instead of the primary antibodies (organ of Corti, the lateral wall, and the spiral ganglion, respectively)

    Article Snippet: BiP/GRP78 and CHOP/Gadd153 Western blot analysis: Higher levels of expression in the experimental groups The results of the Western blot analysis examining the levels of BiP/GRP78 and CHOP/Gadd153 protein are shown in .

    Techniques: Staining, Expressing, Negative Control, Incubation

    Expression of BiP/GRP78 and CHOP/Gadd153 proteins evaluated by Western blot analysis (a). Relative protein level of BiP/GRP 78 and CHOP/Gadd153 was analyzed by using one-way ANOVA (b, c). *Comparison with control group, P

    Journal: Noise & Health

    Article Title: The Protective Effect of the Endoplasmic Reticulum Stress-Related Factors BiP/GRP78 and CHOP/Gadd153 on Noise-induced Hearing Loss in Guinea Pigs

    doi: 10.4103/1463-1741.192481

    Figure Lengend Snippet: Expression of BiP/GRP78 and CHOP/Gadd153 proteins evaluated by Western blot analysis (a). Relative protein level of BiP/GRP 78 and CHOP/Gadd153 was analyzed by using one-way ANOVA (b, c). *Comparison with control group, P

    Article Snippet: BiP/GRP78 and CHOP/Gadd153 Western blot analysis: Higher levels of expression in the experimental groups The results of the Western blot analysis examining the levels of BiP/GRP78 and CHOP/Gadd153 protein are shown in .

    Techniques: Expressing, Western Blot

    Grp78/BiP protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Activation of the endoplasmic reticulum stress response in skeletal muscle of G93A*SOD1 amyotrophic lateral sclerosis mice

    doi: 10.3389/fncel.2015.00170

    Figure Lengend Snippet: Grp78/BiP protein identification and quantitation using label-free spectral counting-based mass spectrometry. (A) Workflow of label-free spectral counting-based protein quantitative analysis using LC-MS/MS. Protein samples were separated using SDS-polyacrylamide gel electrophoresis (PAGE) and gel pieces excised at ~80 kDa for the purpose of in-gel trypsin digestion and LC-MS/MS analysis. Protein quantitative data analysis was conducted using spectral counting and interpreted by normalized total spectra numbers. (B) Grp78/BiP protein identification and peptide coverage using LC-MS/MS. (C) Representative mass-to-charge ratio spectrum and b-/y ions fragmentation of Grp78/BiP peptide (highlighted in B ). (D) Protein quantitative data analysis using the ratio of normalized total spectra numbers of Grp78/BiP to a house keeping protein GAPDH. Three independent muscles from 120–140d old wild type (WT) and ALS-Tg mice were analyzed. * p

    Article Snippet: Samples were then transferred to PVDF membrane (Millipore) and blocked with 5% (w/v) non-fat dry milk in Tris-buffered saline (pH 8.0) for 1 h. The appropriate primary antibodies were added: Grp78/BiP (BD Biosciences) (PERK, phospho-PERK (Thr980), IRE1α, eIF2α, phospho-eIF2α (Ser51), PDI, and CHOP; 1:1000, Cell Signaling Technology) and membranes were incubated at 4°C overnight, washed and then and subsequently probed with HRP-linked anti-rabbit IgG or anti-mouse IgG antibodies (1:1000, Cell Signaling Technology) 1 h at room temperature.

    Techniques: Quantitation Assay, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Polyacrylamide Gel Electrophoresis, Mouse Assay

    Grp78/BiP and CHOP protein levels in cardiac muscle and liver tissue of G93A*SOD1 ALS-Tg mice. (A) Cardiac muscle (HRT) was collected and protein levels determined using western blot technique. Grp78/BiP and CHOP antibodies were used and three postnatal ages were examined: early pre-symptomatic (70d; n = 3 each for WT and ALS-Tg), late pre-symptomatic (90d; n = 5 each for WT and ALS-Tg), and symptomatic (120–140d; n = 3 each for WT and ALS-Tg) mice. (B) Liver tissues (LIV) was collected and protein levels were determined as described above. Analysis of average arbitrary units (AU) obtained by densitometry of Grp78/BiP and CHOP in HRT (C) and in LIV (D) . Data in (C,D) are presented as mean ± S.E.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Activation of the endoplasmic reticulum stress response in skeletal muscle of G93A*SOD1 amyotrophic lateral sclerosis mice

    doi: 10.3389/fncel.2015.00170

    Figure Lengend Snippet: Grp78/BiP and CHOP protein levels in cardiac muscle and liver tissue of G93A*SOD1 ALS-Tg mice. (A) Cardiac muscle (HRT) was collected and protein levels determined using western blot technique. Grp78/BiP and CHOP antibodies were used and three postnatal ages were examined: early pre-symptomatic (70d; n = 3 each for WT and ALS-Tg), late pre-symptomatic (90d; n = 5 each for WT and ALS-Tg), and symptomatic (120–140d; n = 3 each for WT and ALS-Tg) mice. (B) Liver tissues (LIV) was collected and protein levels were determined as described above. Analysis of average arbitrary units (AU) obtained by densitometry of Grp78/BiP and CHOP in HRT (C) and in LIV (D) . Data in (C,D) are presented as mean ± S.E.

    Article Snippet: Samples were then transferred to PVDF membrane (Millipore) and blocked with 5% (w/v) non-fat dry milk in Tris-buffered saline (pH 8.0) for 1 h. The appropriate primary antibodies were added: Grp78/BiP (BD Biosciences) (PERK, phospho-PERK (Thr980), IRE1α, eIF2α, phospho-eIF2α (Ser51), PDI, and CHOP; 1:1000, Cell Signaling Technology) and membranes were incubated at 4°C overnight, washed and then and subsequently probed with HRP-linked anti-rabbit IgG or anti-mouse IgG antibodies (1:1000, Cell Signaling Technology) 1 h at room temperature.

    Techniques: Mouse Assay, Western Blot

    Comparison of Grp78/BiP and CHOP protein levels between white and red gastrocnemius (RG) muscle tissues of G93A*SOD1 ALS-Tg mice. (A) Image of deep portion of gastrocnemius muscle showing white and RG (yellow dashed areas) muscle region. (B) White (WG) and red (RG) gastrocnemius muscle tissues of symptomatic animals (120–140d; n = 3 each for WT and ALS-Tg) were collected. Grp78/BiP and CHOP protein levels were determined using western blot technique. (C) Analysis of Grp78/BiP and CHOP average ratio of ALS-Tg to WT. Data in (B) is presented as mean ± S.E; *, p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Activation of the endoplasmic reticulum stress response in skeletal muscle of G93A*SOD1 amyotrophic lateral sclerosis mice

    doi: 10.3389/fncel.2015.00170

    Figure Lengend Snippet: Comparison of Grp78/BiP and CHOP protein levels between white and red gastrocnemius (RG) muscle tissues of G93A*SOD1 ALS-Tg mice. (A) Image of deep portion of gastrocnemius muscle showing white and RG (yellow dashed areas) muscle region. (B) White (WG) and red (RG) gastrocnemius muscle tissues of symptomatic animals (120–140d; n = 3 each for WT and ALS-Tg) were collected. Grp78/BiP and CHOP protein levels were determined using western blot technique. (C) Analysis of Grp78/BiP and CHOP average ratio of ALS-Tg to WT. Data in (B) is presented as mean ± S.E; *, p

    Article Snippet: Samples were then transferred to PVDF membrane (Millipore) and blocked with 5% (w/v) non-fat dry milk in Tris-buffered saline (pH 8.0) for 1 h. The appropriate primary antibodies were added: Grp78/BiP (BD Biosciences) (PERK, phospho-PERK (Thr980), IRE1α, eIF2α, phospho-eIF2α (Ser51), PDI, and CHOP; 1:1000, Cell Signaling Technology) and membranes were incubated at 4°C overnight, washed and then and subsequently probed with HRP-linked anti-rabbit IgG or anti-mouse IgG antibodies (1:1000, Cell Signaling Technology) 1 h at room temperature.

    Techniques: Mouse Assay, Western Blot

    Schematic figure showing unfolded protein response and endoplasmic reticulum (ER) stress pathway and its proposed role in skeletal muscle atrophy and weakness in ALS . In skeletal muscle of ALS-Tg mice, the G93A*SOD1 mutation leads to oxidative stress and protein misfolding. This leads to an age-dependent activation of ER stress sensors protein kinase RNA-activated-like ER kinase (PERK) and inositol-requiring kinase 1-alpha (IRE1α). Normally, these ER stress sensors physically interact with the ER chaperone immunoglobulin binding protein (Grp78/BiP) which suppresses their activation but accumulation of unfolded/misfolded proteins activates Grp78/BiP, including an upregulation of Grp78/BiP and PDI protein expression. Prolonged and severe ER stress can trigger apoptosis by ER stress-specific cell death signals, including C/EBP homologous protein (CHOP) and caspase-12, leading to muscle atrophy.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Activation of the endoplasmic reticulum stress response in skeletal muscle of G93A*SOD1 amyotrophic lateral sclerosis mice

    doi: 10.3389/fncel.2015.00170

    Figure Lengend Snippet: Schematic figure showing unfolded protein response and endoplasmic reticulum (ER) stress pathway and its proposed role in skeletal muscle atrophy and weakness in ALS . In skeletal muscle of ALS-Tg mice, the G93A*SOD1 mutation leads to oxidative stress and protein misfolding. This leads to an age-dependent activation of ER stress sensors protein kinase RNA-activated-like ER kinase (PERK) and inositol-requiring kinase 1-alpha (IRE1α). Normally, these ER stress sensors physically interact with the ER chaperone immunoglobulin binding protein (Grp78/BiP) which suppresses their activation but accumulation of unfolded/misfolded proteins activates Grp78/BiP, including an upregulation of Grp78/BiP and PDI protein expression. Prolonged and severe ER stress can trigger apoptosis by ER stress-specific cell death signals, including C/EBP homologous protein (CHOP) and caspase-12, leading to muscle atrophy.

    Article Snippet: Samples were then transferred to PVDF membrane (Millipore) and blocked with 5% (w/v) non-fat dry milk in Tris-buffered saline (pH 8.0) for 1 h. The appropriate primary antibodies were added: Grp78/BiP (BD Biosciences) (PERK, phospho-PERK (Thr980), IRE1α, eIF2α, phospho-eIF2α (Ser51), PDI, and CHOP; 1:1000, Cell Signaling Technology) and membranes were incubated at 4°C overnight, washed and then and subsequently probed with HRP-linked anti-rabbit IgG or anti-mouse IgG antibodies (1:1000, Cell Signaling Technology) 1 h at room temperature.

    Techniques: Mouse Assay, Mutagenesis, Activation Assay, Binding Assay, Expressing

    ER chaperones Grp78/BiP and protein disulfide isomerase (PDI) are up-regulated in skeletal muscle of G93A*SOD1 ALS-Tg mice. (A) WG muscle tissues were used to determine Grp78/BiP and PDI expressions using western blot technique from different ages of wild-type (WT) and transgenic G93A*SOD1 (ALS-Tg) mice. Representative images of Grp78/BiP and PDI are shown. Three postnatal ages were examined as follows: early pre-symptomatic (70d; n = 3 each for WT and ALS-Tg), late pre-symptomatic (90d; n = 5 each for WT and ALS-Tg), and symptomatic (120–140d; n = 3 each for WT and ALS-Tg) mice. (B,C) Analysis of average arbitrary units (AU) obtained by densitometry of PDI. Data in B are presented as mean ± S.E; **, p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Activation of the endoplasmic reticulum stress response in skeletal muscle of G93A*SOD1 amyotrophic lateral sclerosis mice

    doi: 10.3389/fncel.2015.00170

    Figure Lengend Snippet: ER chaperones Grp78/BiP and protein disulfide isomerase (PDI) are up-regulated in skeletal muscle of G93A*SOD1 ALS-Tg mice. (A) WG muscle tissues were used to determine Grp78/BiP and PDI expressions using western blot technique from different ages of wild-type (WT) and transgenic G93A*SOD1 (ALS-Tg) mice. Representative images of Grp78/BiP and PDI are shown. Three postnatal ages were examined as follows: early pre-symptomatic (70d; n = 3 each for WT and ALS-Tg), late pre-symptomatic (90d; n = 5 each for WT and ALS-Tg), and symptomatic (120–140d; n = 3 each for WT and ALS-Tg) mice. (B,C) Analysis of average arbitrary units (AU) obtained by densitometry of PDI. Data in B are presented as mean ± S.E; **, p

    Article Snippet: Samples were then transferred to PVDF membrane (Millipore) and blocked with 5% (w/v) non-fat dry milk in Tris-buffered saline (pH 8.0) for 1 h. The appropriate primary antibodies were added: Grp78/BiP (BD Biosciences) (PERK, phospho-PERK (Thr980), IRE1α, eIF2α, phospho-eIF2α (Ser51), PDI, and CHOP; 1:1000, Cell Signaling Technology) and membranes were incubated at 4°C overnight, washed and then and subsequently probed with HRP-linked anti-rabbit IgG or anti-mouse IgG antibodies (1:1000, Cell Signaling Technology) 1 h at room temperature.

    Techniques: Mouse Assay, Western Blot, Transgenic Assay