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  • 92
    Santa Cruz Biotechnology grp78
    CDK1 or CDK5 are required for XBP-1s and <t>Grp78</t> induction (A) U937, K562 cells were transfected with shCDK1/pLKO.1 or shControl/pLKO.1 for 48h, after which cells were exposed to Tg (1-5μM) for 3-6h, followed by Western blot analysis. (B) J558 cells were treated with RO3306 (5-15 μM), AZD5438 (0.2-4 μM) or JNJ7706621 (0.2-4 μM) for 6h, after which Western blot analysis was performed to monitor XBP-1s expression. (C) K562 or Jurkat cells were transfected with shCDK5/pLKO.1 or shCont for 48h, followed by exposure to Tg (1.5-5 μM) for 4h, after which cell extracts were subjected to WB analysis using the indicated primary antibodies. (D) J558 cells were treated with PHA793887 (0.02-5 μM) for 6h, after which Western blot analysis was performed to monitor XBP-1s expression.
    Grp78, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 2238 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    grp78  (Abcam)
    92
    Abcam grp78
    a BrdU labelling of control primary chondrocytes and chondrocytes treated with tunicamycin with and without exogenous MANF ( n = 2). b Western blot and densitometry measurement of <t>GRP78</t> levels in untreated and tunicamycin-treated primary chondrocytes with and without exogenous MANF ( n = 2). Tun tunicamycin. *** P
    Grp78, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 1424 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc grp78
    Effect of dicerandrol B on ERS in HeLa cells. Notes: ( A – C ) Western blot analysis of <t>GRP78</t> and Ub proteins in HeLa cells incubated with dicerandrol B (0, 3, or 5 µg/mL) for 12 hours. Data are presented as mean ± SD of three experiments. (* P
    Grp78, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1227 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology anti grp78
    Protein expression levels of apoptotic and ER stress markers following H 2 O 2 treatment of vector, NDRG2 and 3A-NDRG2-infected C2C12 myoblasts at P3. Protein expression levels of (A) total and cleaved PARP, (B) total and cleaved caspase 3, (C) Bcl-2, (D) Bcl-xL, (E) Mcl-1, (F) Bax, (G) <t>GRP78</t> and (H) NDRG2 proteins. Alpha-tubulin (α-Tubulin) protein indicates protein levels loaded. Hydrogen peroxide treatment times were 0 h (black bars), 4 h (white bars) or 8 h (gray bars). Data are the mean of three independent experiments ( n = 3 per treatment). *** P
    Anti Grp78, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 680 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti grp78
    Increased <t>GRP78</t> expression (6A), GRP94 expression (6B) and caspase 12 activation in anoxia-reoxygenation induced ERS response were abrogated by adding levosimendan, each result obtained from sample of 3×10 6 cells. Each point represents mean ± S.D. of three independent experiments. * p
    Anti Grp78, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 544 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson grp78
    scRNAseq of peripheral lung tissue in nonfibrotic individuals. Violin plots for expression levels of A: ACE2 , B: TMPRSS2 , C: ADAM17 , D: CTSL (Cathepsin L) , E: CD147 , and F: <t>GRP78</t> across lung epithelial cell populations in healthy subjects (see methods for dataset reference for cell population markers).
    Grp78, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 234 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti grp78 bip antibody
    Estradiol treatment upregulates the expression of endoplasmic reticulum stress (ERS) related proteins in esophageal cancer cell line EC109. The cells were cultured in serum-free Roswell Park Memorial Institute (RPMI) 1640 medium for 24 h, and treated with 17-estradiol (E2), E2 and ICI 182,780 (E2 + ICI), E2 and 4-phenylbutyric acid (4-PBA), and tunicamycin (TM) for 24 h, respectively. (A) <t>GRP78</t> proteins in EC109 cells under an Olympus laser confocal microscope. (B–E) The expressions of GRP78, ATF6, IRE1α, and PERK in EC109 cells after various treatments. Densitometric values were normalized to β-actin. The results are expressed as the means ± SEM, n = 3. * P
    Anti Grp78 Bip Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc anti grp78
    LRRK2 supports <t>GRP78-mediated</t> cell survival. ( A ) Quantification of the populations of cells with sub-G0 DNA content in cultures of SH-SY5Y cells transfected with a control vector (Control) and MIX LRRK2 KD SH-SY5Y cells (LRRK2 KD) treated with 0, 3 or 6 µM tunicamycin for 16 hours. Data represent the mean ± SEM of 3 independent experiments. ***, p
    Anti Grp78, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 303 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher grp78
    Effects of LM-1685, a non-coxib COX-2 inhibitor, on the viability and induction of ER stress-related molecules in NTUB1 and T24 cells. (A) Cells were treated with LM-1685 (0–160 µM) for 24 h. Cell viability was analyzed by MTT assay. Data are presented as means ± SD of three independents experiments. (B) Cells were treated with LM-1685 (80 and 160 µM) or celecoxib (100 µM) for 24 h. The cell lysates were harvested at each time point and analyzed by Western blotting with specific antibodies to detect ER stress-related molecules IRE-1α, CHOP, calnexin, and <t>GRP78,</t> and caspase-4. CF is the abbreviation of cleaved form. (C). Cells were transfected with GRP78 siRNA (10 nM) or scramble siRNA (10 nM) (as a control), and then treated with 160 µM LM-1685. The combinative effect of LM-1685 and GRP78 knockdown on apoptosis was determined by flow cytometry (FACS) with annexin V-FITC and PI labeling. The quantitative analysis of total apoptosis (early and late) population was presented. Data are presented as means ± SD of three independents experiments. * p
    Grp78, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 261 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti grp78
    Effects of darbepoetin alfa on left ventricular myocardial <t>GRP78,</t> CHOP, ATF6, and caspase-12 in the Sham– and β 1 -EC II –immunized rabbits. Representative immunoblots are shown on the left to the bar graphs. N=6 animals in each group. Bar denote SEM. *P
    Anti Grp78, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 167 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    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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    92
    Bioneer Corporation grp78
    HCV E2 inhibited the interaction between <t>grp78</t> and AIMP1/p43. A. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody. B. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells along with 2 or 4 µg of plasmid expressing HCV E2 and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody. C. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells along with plasmid expressing HCV E2, E1, core, or NS5A and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody.
    Grp78, supplied by Bioneer Corporation, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam rabbit anti grp78
    Semi-quantitative analysis of immunohistochemical staining of <t>GRP78,</t> p-ERK and caspase-12 in the kidney, based on the percentage of positively stained areas in the tubules and the intensity of staining. Animals were divided into four groups: the control group (N), the cisplatin (CP) group (C), the grape seed proanthocyanidin extract (GSPE) group (G) and the CP+GSPE group (C+G). The expression of GRP78, p-ERK and caspase-12 was significantly increased in the C group compared to the N group and significantly decreased in the C+G group compared to the C group. * Statistically significant difference compared to the N group (p
    Rabbit Anti Grp78, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 102 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    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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    Proteintech rabbit grp78 bip polyclonal
    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 .
    Rabbit Grp78 Bip Polyclonal, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 83 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Becton Dickinson anti bip grp78
    Synaptic markers in VD. Pre-synaptic, pos-synaptic and mitochondrial proteins were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of VD patients and controls. The levels of synaptophysin, PSD95 and HSP60 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of <t>GRP78</t> and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, *p
    Anti Bip Grp78, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    CDK1 or CDK5 are required for XBP-1s and Grp78 induction (A) U937, K562 cells were transfected with shCDK1/pLKO.1 or shControl/pLKO.1 for 48h, after which cells were exposed to Tg (1-5μM) for 3-6h, followed by Western blot analysis. (B) J558 cells were treated with RO3306 (5-15 μM), AZD5438 (0.2-4 μM) or JNJ7706621 (0.2-4 μM) for 6h, after which Western blot analysis was performed to monitor XBP-1s expression. (C) K562 or Jurkat cells were transfected with shCDK5/pLKO.1 or shCont for 48h, followed by exposure to Tg (1.5-5 μM) for 4h, after which cell extracts were subjected to WB analysis using the indicated primary antibodies. (D) J558 cells were treated with PHA793887 (0.02-5 μM) for 6h, after which Western blot analysis was performed to monitor XBP-1s expression.

    Journal: Molecular cancer therapeutics

    Article Title: DINACICLIB (SCH727965) INHIBITS THE UNFOLDED PROTEIN RESPONSE (UPR) THROUGH A CDK1 AND CDK5-DEPENDENT MECHANISM

    doi: 10.1158/1535-7163.MCT-13-0714

    Figure Lengend Snippet: CDK1 or CDK5 are required for XBP-1s and Grp78 induction (A) U937, K562 cells were transfected with shCDK1/pLKO.1 or shControl/pLKO.1 for 48h, after which cells were exposed to Tg (1-5μM) for 3-6h, followed by Western blot analysis. (B) J558 cells were treated with RO3306 (5-15 μM), AZD5438 (0.2-4 μM) or JNJ7706621 (0.2-4 μM) for 6h, after which Western blot analysis was performed to monitor XBP-1s expression. (C) K562 or Jurkat cells were transfected with shCDK5/pLKO.1 or shCont for 48h, followed by exposure to Tg (1.5-5 μM) for 4h, after which cell extracts were subjected to WB analysis using the indicated primary antibodies. (D) J558 cells were treated with PHA793887 (0.02-5 μM) for 6h, after which Western blot analysis was performed to monitor XBP-1s expression.

    Article Snippet: Finally, J558 cells treated with a specific CDK4/6 inhibitor (PD332991) , or a CDK 9/2/7 inhibitor (SNS-032) ( ) did not display changes in Grp78 or XBP-1s expression (data not shown).

    Techniques: Transfection, Western Blot, Expressing

    Significant upregulation of anti-apoptotic factors, GRP 78 and SYVN1, in 2-month-old 5×FAD mice. (a) GRP 78 expression of 5×FAD mice and age-matched WT mice at 2, 7, and 12 months old ( n = 6, * P

    Journal: Chinese Medical Journal

    Article Title: Endoplasmic Reticulum Stress Induces the Early Appearance of Pro-apoptotic and Anti-apoptotic Proteins in Neurons of Five Familial Alzheimer's Disease Mice

    doi: 10.4103/0366-6999.194643

    Figure Lengend Snippet: Significant upregulation of anti-apoptotic factors, GRP 78 and SYVN1, in 2-month-old 5×FAD mice. (a) GRP 78 expression of 5×FAD mice and age-matched WT mice at 2, 7, and 12 months old ( n = 6, * P

    Article Snippet: Our results showed that the expression of GRP 78 and SYVN1 changed without statistical significance but displayed an early-increase and subsequent-decline tendency from 2 to 12 months in WT mice.

    Techniques: Mouse Assay, Expressing

    Schematic drawing shows that activated UPR pathways of ERS induced by intraneuronal β-amyloid in 5×FAD mice. Aβ in 2-month-old 5×FAD brains lead to higher ERS level than that of WT mice, which produces a cellular protective effect, on the one hand, up-regulating the expression of ER chaperones and related degradation proteins mainly via p-IRE-1α-XBP1s and ATF6α pathways at the early stage of AD to eliminate Aβ; on the other hand, inducing increased expression of downstream signaling pathways molecules, especially pro-apoptotic proteins (cleaved caspase-12 and CHOP). Under sustained stress conditions caused by the Aβ-associated toxic effects, the protective function of the UPR declines and the pro-apoptotic functions are enhanced gradually. The pro-apoptotic functions of the UPR may lead to functional impairment of neurons and even death. UPR: Unfolded protein response; ERS: Endoplasmic reticulum stress; Aβ: Amyloid β; 5×FAD: Transgenic mice with five familiar Alzheimer's disease; p-PERK: Phosphorylated protein kinase RNA-like ER kinase; p-eIF2α: Phosphorylated eukaryotic translation initiation factor 2α; ATF4: Activating transcription factor 4; CHOP: CCAAT/enhancer-binding protein homologous protein; p-IRE-1α: Phosphorylated inositol-requiring enzyme 1α; XBP1s: Spliced X box-binding protein 1; ATF6α: Activating transcription factor 6α; GRP 78: Glucose-regulated protein 78; ERAD: Endoplasmic reticulum-associated protein degradation; SYVN1: Ubiquitin ligase synovial apoptosis inhibitor 1; TRAF2: Tumor necrosis factor-receptor-associated factor 2; ASK1: Apoptosis signal-regulating kinase 1; JNK: c-Jun amino-terminal kinase.

    Journal: Chinese Medical Journal

    Article Title: Endoplasmic Reticulum Stress Induces the Early Appearance of Pro-apoptotic and Anti-apoptotic Proteins in Neurons of Five Familial Alzheimer's Disease Mice

    doi: 10.4103/0366-6999.194643

    Figure Lengend Snippet: Schematic drawing shows that activated UPR pathways of ERS induced by intraneuronal β-amyloid in 5×FAD mice. Aβ in 2-month-old 5×FAD brains lead to higher ERS level than that of WT mice, which produces a cellular protective effect, on the one hand, up-regulating the expression of ER chaperones and related degradation proteins mainly via p-IRE-1α-XBP1s and ATF6α pathways at the early stage of AD to eliminate Aβ; on the other hand, inducing increased expression of downstream signaling pathways molecules, especially pro-apoptotic proteins (cleaved caspase-12 and CHOP). Under sustained stress conditions caused by the Aβ-associated toxic effects, the protective function of the UPR declines and the pro-apoptotic functions are enhanced gradually. The pro-apoptotic functions of the UPR may lead to functional impairment of neurons and even death. UPR: Unfolded protein response; ERS: Endoplasmic reticulum stress; Aβ: Amyloid β; 5×FAD: Transgenic mice with five familiar Alzheimer's disease; p-PERK: Phosphorylated protein kinase RNA-like ER kinase; p-eIF2α: Phosphorylated eukaryotic translation initiation factor 2α; ATF4: Activating transcription factor 4; CHOP: CCAAT/enhancer-binding protein homologous protein; p-IRE-1α: Phosphorylated inositol-requiring enzyme 1α; XBP1s: Spliced X box-binding protein 1; ATF6α: Activating transcription factor 6α; GRP 78: Glucose-regulated protein 78; ERAD: Endoplasmic reticulum-associated protein degradation; SYVN1: Ubiquitin ligase synovial apoptosis inhibitor 1; TRAF2: Tumor necrosis factor-receptor-associated factor 2; ASK1: Apoptosis signal-regulating kinase 1; JNK: c-Jun amino-terminal kinase.

    Article Snippet: Our results showed that the expression of GRP 78 and SYVN1 changed without statistical significance but displayed an early-increase and subsequent-decline tendency from 2 to 12 months in WT mice.

    Techniques: Mouse Assay, Expressing, Functional Assay, Transgenic Assay, Binding Assay

    a BrdU labelling of control primary chondrocytes and chondrocytes treated with tunicamycin with and without exogenous MANF ( n = 2). b Western blot and densitometry measurement of GRP78 levels in untreated and tunicamycin-treated primary chondrocytes with and without exogenous MANF ( n = 2). Tun tunicamycin. *** P

    Journal: Cell Stress & Chaperones

    Article Title: Mesencephalic astrocyte-derived neurotropic factor is an important factor in chondrocyte ER homeostasis

    doi: 10.1007/s12192-018-0953-7

    Figure Lengend Snippet: a BrdU labelling of control primary chondrocytes and chondrocytes treated with tunicamycin with and without exogenous MANF ( n = 2). b Western blot and densitometry measurement of GRP78 levels in untreated and tunicamycin-treated primary chondrocytes with and without exogenous MANF ( n = 2). Tun tunicamycin. *** P

    Article Snippet: However, in vitro treatment of Matn3 V194D chondrocytes with exogenous MANF had no effect on the amount or localisation of retained mutant matrilin-3 despite lowering the levels of GRP78 by 25% (Fig. b–d).

    Techniques: Western Blot

    a X-ray radiographs of Manf fl/fl Col2Cre − Matn3 +/+ , Manf fl/fl Col2Cre − Matn3 V194D/V194D and Manf fl/fl Col2Cre + Matn3 V194D/V194D mice at 6 weeks, showing the dramatic effect of removal of MANF from cartilage of the mouse model of MED. b Western blot and densitometry measurement of intracellular GRP78 levels in Matn3 V194D/V194D primary chondrocytes with and without exogenous MANF ( n = 4). c Western blot and densitometry measurement of intracellular matrilin-3 levels in Matn3 V194D/V194D primary chondrocytes with and without exogenous MANF ( n = 4). d Immunocytochemistry for matrilin-3 showing that exogenous MANF has no effect on the intracellular retention of mutant matrilin-3 (in green) in Matn3 V194D/V194D primary chondrocytes. DAPI was used as a counterstain. Scale bars 5 mm ( a ) and 200 μm ( d ) (colour figure online)

    Journal: Cell Stress & Chaperones

    Article Title: Mesencephalic astrocyte-derived neurotropic factor is an important factor in chondrocyte ER homeostasis

    doi: 10.1007/s12192-018-0953-7

    Figure Lengend Snippet: a X-ray radiographs of Manf fl/fl Col2Cre − Matn3 +/+ , Manf fl/fl Col2Cre − Matn3 V194D/V194D and Manf fl/fl Col2Cre + Matn3 V194D/V194D mice at 6 weeks, showing the dramatic effect of removal of MANF from cartilage of the mouse model of MED. b Western blot and densitometry measurement of intracellular GRP78 levels in Matn3 V194D/V194D primary chondrocytes with and without exogenous MANF ( n = 4). c Western blot and densitometry measurement of intracellular matrilin-3 levels in Matn3 V194D/V194D primary chondrocytes with and without exogenous MANF ( n = 4). d Immunocytochemistry for matrilin-3 showing that exogenous MANF has no effect on the intracellular retention of mutant matrilin-3 (in green) in Matn3 V194D/V194D primary chondrocytes. DAPI was used as a counterstain. Scale bars 5 mm ( a ) and 200 μm ( d ) (colour figure online)

    Article Snippet: However, in vitro treatment of Matn3 V194D chondrocytes with exogenous MANF had no effect on the amount or localisation of retained mutant matrilin-3 despite lowering the levels of GRP78 by 25% (Fig. b–d).

    Techniques: Mouse Assay, Western Blot, Immunocytochemistry, Mutagenesis

    a STRING network showing the known interactions between the proteins encoded by the genes differentially expressed in the Manf fl/fl Col2Cre − and Col2Cre + cartilage at P5. b Deletion of MANF increases the levels of GRP78 but not GRP94 in cartilage at P5, as shown by Western blotting and densitometry measurement ( n = 3). c Western blotting of proteins co-immunoprecipitated in the 293 cells expressing FLAG-tagged recombinant MANF

    Journal: Cell Stress & Chaperones

    Article Title: Mesencephalic astrocyte-derived neurotropic factor is an important factor in chondrocyte ER homeostasis

    doi: 10.1007/s12192-018-0953-7

    Figure Lengend Snippet: a STRING network showing the known interactions between the proteins encoded by the genes differentially expressed in the Manf fl/fl Col2Cre − and Col2Cre + cartilage at P5. b Deletion of MANF increases the levels of GRP78 but not GRP94 in cartilage at P5, as shown by Western blotting and densitometry measurement ( n = 3). c Western blotting of proteins co-immunoprecipitated in the 293 cells expressing FLAG-tagged recombinant MANF

    Article Snippet: However, in vitro treatment of Matn3 V194D chondrocytes with exogenous MANF had no effect on the amount or localisation of retained mutant matrilin-3 despite lowering the levels of GRP78 by 25% (Fig. b–d).

    Techniques: Western Blot, Immunoprecipitation, Expressing, Recombinant

    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

    NIM811 induced a transient upregulation of UPR and autophagy. ( a and b ) In all, 0 μ M NIM811 transiently induced UPR signaling (P-EIF2a, Bip, ATF4, CHOP upregulation) at 6 h, but except for Bip, the activation of UPR mediators disappeared at 9 h. Tunicamycin (Tm) and thapsigargin (Tg) were used as positive controls for the UPR. ( c ) In total, 10 μ M NIM811 was added to GFP-LC3B transfected U251 cells and imaged by fluorescence microscopy at 0 h and 6 h. Scale bar=25 μ m. LC3 puncta were detected at 3–6 h. LC3 puncta formation occurred before the appearance of vacuoles. ( d ) LC3-I and -II conversion occurred at 4–6 h of 10 μ M NIM811 treatment accompanied by p62 degradation. ( e and f ) In all, 10 μ M NIM811 incubation caused p62 accumulation at 24 h, which persisted at later time points (40–46 h). ( g ) Immunofluorescence staining of SQSTM1/p62 (red) and nucleus (blue) after 24 h vehicle (left) or 10 μ M NIM811-treated (right) U251 cells. Scale bar=25 μ m

    Journal: Cell Death & Disease

    Article Title: Cycloheximide promotes paraptosis induced by inhibition of cyclophilins in glioblastoma multiforme

    doi: 10.1038/cddis.2017.217

    Figure Lengend Snippet: NIM811 induced a transient upregulation of UPR and autophagy. ( a and b ) In all, 0 μ M NIM811 transiently induced UPR signaling (P-EIF2a, Bip, ATF4, CHOP upregulation) at 6 h, but except for Bip, the activation of UPR mediators disappeared at 9 h. Tunicamycin (Tm) and thapsigargin (Tg) were used as positive controls for the UPR. ( c ) In total, 10 μ M NIM811 was added to GFP-LC3B transfected U251 cells and imaged by fluorescence microscopy at 0 h and 6 h. Scale bar=25 μ m. LC3 puncta were detected at 3–6 h. LC3 puncta formation occurred before the appearance of vacuoles. ( d ) LC3-I and -II conversion occurred at 4–6 h of 10 μ M NIM811 treatment accompanied by p62 degradation. ( e and f ) In all, 10 μ M NIM811 incubation caused p62 accumulation at 24 h, which persisted at later time points (40–46 h). ( g ) Immunofluorescence staining of SQSTM1/p62 (red) and nucleus (blue) after 24 h vehicle (left) or 10 μ M NIM811-treated (right) U251 cells. Scale bar=25 μ m

    Article Snippet: Antibodies used were the followings: mouse anti-ubiquitin (13-1600, Invitrogen, Camarillo, CA, USA), mouse actin (mab1501, EMD Millipore, Billerica, MA, USA), ATF4 (sc-200, Santa Cruz Biotechnology, Dallas, TX, USA), EIF2a (#9722, Cell Signaling Technology, Danvers, MA, USA), P-EIF2a (#9721, Cell Signaling Technology), Bip (ab21685, Abcam, Cambridge, UK), p62 (ab194129, Abcam), LC3B (NB100-2220, Novus Biologicals, Littleton, CO, USA), P-mTOR (#2971, Cell Signaling Technology), P-AKT (#9271, Cell Signaling Technology) and P-P70S6K (#9205, Cell Signaling Technology).

    Techniques: Activation Assay, Transfection, Fluorescence, Microscopy, Incubation, Immunofluorescence, Staining

    Effect of dicerandrol B on ERS in HeLa cells. Notes: ( A – C ) Western blot analysis of GRP78 and Ub proteins in HeLa cells incubated with dicerandrol B (0, 3, or 5 µg/mL) for 12 hours. Data are presented as mean ± SD of three experiments. (* P

    Journal: OncoTargets and therapy

    Article Title: Dicerandrol B: a natural xanthone dimer induces apoptosis in cervical cancer HeLa cells through the endoplasmic reticulum stress and mitochondrial damage

    doi: 10.2147/OTT.S191204

    Figure Lengend Snippet: Effect of dicerandrol B on ERS in HeLa cells. Notes: ( A – C ) Western blot analysis of GRP78 and Ub proteins in HeLa cells incubated with dicerandrol B (0, 3, or 5 µg/mL) for 12 hours. Data are presented as mean ± SD of three experiments. (* P

    Article Snippet: GRP78 is a major ER chaperone that exerts anti-apoptotic effects and regulates the ERS response., The mitochondrial pathway is another important apop-totic pathway.

    Techniques: Western Blot, Incubation

    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

    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

    Protein expression levels of apoptotic and ER stress markers following H 2 O 2 treatment of vector, NDRG2 and 3A-NDRG2-infected C2C12 myoblasts at P3. Protein expression levels of (A) total and cleaved PARP, (B) total and cleaved caspase 3, (C) Bcl-2, (D) Bcl-xL, (E) Mcl-1, (F) Bax, (G) GRP78 and (H) NDRG2 proteins. Alpha-tubulin (α-Tubulin) protein indicates protein levels loaded. Hydrogen peroxide treatment times were 0 h (black bars), 4 h (white bars) or 8 h (gray bars). Data are the mean of three independent experiments ( n = 3 per treatment). *** P

    Journal: FEBS Open Bio

    Article Title: NDRG2 promotes myoblast proliferation and caspase 3/7 activities during differentiation, and attenuates hydrogen peroxide – But not palmitate-induced toxicity

    doi: 10.1016/j.fob.2015.08.001

    Figure Lengend Snippet: Protein expression levels of apoptotic and ER stress markers following H 2 O 2 treatment of vector, NDRG2 and 3A-NDRG2-infected C2C12 myoblasts at P3. Protein expression levels of (A) total and cleaved PARP, (B) total and cleaved caspase 3, (C) Bcl-2, (D) Bcl-xL, (E) Mcl-1, (F) Bax, (G) GRP78 and (H) NDRG2 proteins. Alpha-tubulin (α-Tubulin) protein indicates protein levels loaded. Hydrogen peroxide treatment times were 0 h (black bars), 4 h (white bars) or 8 h (gray bars). Data are the mean of three independent experiments ( n = 3 per treatment). *** P

    Article Snippet: Rabbit polyclonal antibodies anti-Myf5 (sc-302; 1:200) and anti-MyoD (sc-760; 1:350), and mouse monoclonal antibodies anti-myogenin (sc-12732; 1:200) and anti-GRP78 (sc-376768; 1:200) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Expressing, Plasmid Preparation, Infection

    GRP78 expression is elevated in highly metastatic tumors and is available for therapeutic targeting

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Inhibition of Established Micrometastases by Targeted Drug Delivery via Cell Surface Associated GRP78

    doi: 10.1158/1078-0432.CCR-12-2991

    Figure Lengend Snippet: GRP78 expression is elevated in highly metastatic tumors and is available for therapeutic targeting

    Article Snippet: It is possible that cells with low surface GRP78 are still killed effectively by BMTP78 and/or that surface GRP78 is internalized rapidly, such that the acute administration of anti-GRP78 antibody as shown in does not detect all positive cells.

    Techniques: Expressing

    GRP78 staining of primary breast carcinomas and their matched metastases

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Inhibition of Established Micrometastases by Targeted Drug Delivery via Cell Surface Associated GRP78

    doi: 10.1158/1078-0432.CCR-12-2991

    Figure Lengend Snippet: GRP78 staining of primary breast carcinomas and their matched metastases

    Article Snippet: It is possible that cells with low surface GRP78 are still killed effectively by BMTP78 and/or that surface GRP78 is internalized rapidly, such that the acute administration of anti-GRP78 antibody as shown in does not detect all positive cells.

    Techniques: Staining

    Atmospheric oxygen concentrations increase surface localized GRP78 in non-tumorigenic cells

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Inhibition of Established Micrometastases by Targeted Drug Delivery via Cell Surface Associated GRP78

    doi: 10.1158/1078-0432.CCR-12-2991

    Figure Lengend Snippet: Atmospheric oxygen concentrations increase surface localized GRP78 in non-tumorigenic cells

    Article Snippet: It is possible that cells with low surface GRP78 are still killed effectively by BMTP78 and/or that surface GRP78 is internalized rapidly, such that the acute administration of anti-GRP78 antibody as shown in does not detect all positive cells.

    Techniques:

    GRP78-mediated suppression of tumor growth and metastasis following BMTP78 treatment

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Inhibition of Established Micrometastases by Targeted Drug Delivery via Cell Surface Associated GRP78

    doi: 10.1158/1078-0432.CCR-12-2991

    Figure Lengend Snippet: GRP78-mediated suppression of tumor growth and metastasis following BMTP78 treatment

    Article Snippet: It is possible that cells with low surface GRP78 are still killed effectively by BMTP78 and/or that surface GRP78 is internalized rapidly, such that the acute administration of anti-GRP78 antibody as shown in does not detect all positive cells.

    Techniques:

    Occurrence of ER stress and the UPR in a subset of A-NEC patients. (A) Splicing of XBP1 mRNA was checked in all patients, and representative PCR products of XBP1u and XBP1s are shown using DNA electrophoresis. Dimethyl sulfoxide (DMSO)-treated HIEC cells were used as negative control for XBP1s , and TM-treated HIEC cells were used as positive control for XBP1s . Mucosal mRNA expression levels of GRP78 (B) and CHOP (C) in the ileum of patients were quantified using qPCR and normalized to GAPDH mRNA levels. Asterisks indicate statistical significant differences between indicated groups. (D) The Western blot result shows the representative protein expression of GRP78 and CHOP in A-NEC-XBP1u and A-NEC-XBP1s patients, and beta ACTIN was used as loading control.

    Journal: PLoS ONE

    Article Title: Endoplasmic Reticulum Stress, Unfolded Protein Response and Altered T Cell Differentiation in Necrotizing Enterocolitis

    doi: 10.1371/journal.pone.0078491

    Figure Lengend Snippet: Occurrence of ER stress and the UPR in a subset of A-NEC patients. (A) Splicing of XBP1 mRNA was checked in all patients, and representative PCR products of XBP1u and XBP1s are shown using DNA electrophoresis. Dimethyl sulfoxide (DMSO)-treated HIEC cells were used as negative control for XBP1s , and TM-treated HIEC cells were used as positive control for XBP1s . Mucosal mRNA expression levels of GRP78 (B) and CHOP (C) in the ileum of patients were quantified using qPCR and normalized to GAPDH mRNA levels. Asterisks indicate statistical significant differences between indicated groups. (D) The Western blot result shows the representative protein expression of GRP78 and CHOP in A-NEC-XBP1u and A-NEC-XBP1s patients, and beta ACTIN was used as loading control.

    Article Snippet: Antibodies against GRP78 (Santa Cruz Biotechnology, CA, USA), human alpha defensin 5 (HD5, HyCult Biotechnology, Uden, the Netherlands), and Forkhead box P3 (FOXP3, eBioscience, San Diego, CA, USA) were used in combination with the Vectastain Elite ABC kit (Vector Laboratories, Burlingame, CA, USA) and 3,3’-diaminobenzidine as staining reagent for IHC, or in combination with DyLight 594 conjugated secondary antibody and DyLight 488 conjugated secondary antibody (Jackson ImmunoResearch, West Grove, PA, USA) for IF.

    Techniques: Polymerase Chain Reaction, Nucleic Acid Electrophoresis, Negative Control, Positive Control, Expressing, Real-time Polymerase Chain Reaction, Western Blot

    Increased GRP78 expression (6A), GRP94 expression (6B) and caspase 12 activation in anoxia-reoxygenation induced ERS response were abrogated by adding levosimendan, each result obtained from sample of 3×10 6 cells. Each point represents mean ± S.D. of three independent experiments. * p

    Journal: PLoS ONE

    Article Title: Inhibition of Reverse-Mode Sodium-Calcium Exchanger Activity and Apoptosis by Levosimendan in Human Cardiomyocyte Progenitor Cell-Derived Cardiomyocytes after Anoxia and Reoxygenation

    doi: 10.1371/journal.pone.0085909

    Figure Lengend Snippet: Increased GRP78 expression (6A), GRP94 expression (6B) and caspase 12 activation in anoxia-reoxygenation induced ERS response were abrogated by adding levosimendan, each result obtained from sample of 3×10 6 cells. Each point represents mean ± S.D. of three independent experiments. * p

    Article Snippet: Anti-GRP78 (1∶2000, Abcam, Cambridge, MA), anti-caspase-12 (1∶2000, Abcam, Cambridge, MA) and anti-β-actin (1∶30000, Sigma, St.Louis, MO) were obtained.

    Techniques: Expressing, Activation Assay

    Effect of levosimendan on NCX activity in cultured human cardiomyocyte progenitor cell (hCPC)-derived cardiomyocytes after anoxia/reoxygenation (A/R). (A) Reverse-mode NCX activity was measured with or without treatment of Levosimendan under control conditions (a), after anoxia for 2 hours (b), and after anoxia for 14 hours (c). Reverse-mode NCX activity 2 and 14 hours of anoxia were determined; higher expression of GRP78 and caspase 12 did not reach statistical significance until 14 hours of anoxia ( Figure 7 ). (B) NCX activity was estimated by amplitude of increase in intracellular calcium concentration. (Δ[Ca 2+ ] i = peak [Ca 2+ ] i - basal [Ca 2+ ] i ). ( n = 15 for each experimental group). Values are expressed as mean ± S.D. from three independent trials. * p

    Journal: PLoS ONE

    Article Title: Inhibition of Reverse-Mode Sodium-Calcium Exchanger Activity and Apoptosis by Levosimendan in Human Cardiomyocyte Progenitor Cell-Derived Cardiomyocytes after Anoxia and Reoxygenation

    doi: 10.1371/journal.pone.0085909

    Figure Lengend Snippet: Effect of levosimendan on NCX activity in cultured human cardiomyocyte progenitor cell (hCPC)-derived cardiomyocytes after anoxia/reoxygenation (A/R). (A) Reverse-mode NCX activity was measured with or without treatment of Levosimendan under control conditions (a), after anoxia for 2 hours (b), and after anoxia for 14 hours (c). Reverse-mode NCX activity 2 and 14 hours of anoxia were determined; higher expression of GRP78 and caspase 12 did not reach statistical significance until 14 hours of anoxia ( Figure 7 ). (B) NCX activity was estimated by amplitude of increase in intracellular calcium concentration. (Δ[Ca 2+ ] i = peak [Ca 2+ ] i - basal [Ca 2+ ] i ). ( n = 15 for each experimental group). Values are expressed as mean ± S.D. from three independent trials. * p

    Article Snippet: Anti-GRP78 (1∶2000, Abcam, Cambridge, MA), anti-caspase-12 (1∶2000, Abcam, Cambridge, MA) and anti-β-actin (1∶30000, Sigma, St.Louis, MO) were obtained.

    Techniques: Activity Assay, Cell Culture, Derivative Assay, Expressing, Concentration Assay

    Effect of musclin on ERS markers in rat soleus muscles. a : Western blot analysis of the protein levels of the 78-kDa glucose-regulated protein (GRP78) and the 94-kDa GRP94 in soleus muscles. b : Quantitative analysis of GRP78 protein expression. c : Quantitative analysis of GRP94 protein expression. Values are the mean ± standard error of the mean; ( n = 3); * P

    Journal: Nutrition & Metabolism

    Article Title: Positive association between musclin and insulin resistance in obesity: evidence of a human study and an animal experiment

    doi: 10.1186/s12986-017-0199-x

    Figure Lengend Snippet: Effect of musclin on ERS markers in rat soleus muscles. a : Western blot analysis of the protein levels of the 78-kDa glucose-regulated protein (GRP78) and the 94-kDa GRP94 in soleus muscles. b : Quantitative analysis of GRP78 protein expression. c : Quantitative analysis of GRP94 protein expression. Values are the mean ± standard error of the mean; ( n = 3); * P

    Article Snippet: Antibodies against GRP78, GRP94 were from Abcam (Cambridge, UK).

    Techniques: Western Blot, Expressing

    scRNAseq of peripheral lung tissue in nonfibrotic individuals. Violin plots for expression levels of A: ACE2 , B: TMPRSS2 , C: ADAM17 , D: CTSL (Cathepsin L) , E: CD147 , and F: GRP78 across lung epithelial cell populations in healthy subjects (see methods for dataset reference for cell population markers).

    Journal: bioRxiv

    Article Title: Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue

    doi: 10.1101/2020.04.07.030742

    Figure Lengend Snippet: scRNAseq of peripheral lung tissue in nonfibrotic individuals. Violin plots for expression levels of A: ACE2 , B: TMPRSS2 , C: ADAM17 , D: CTSL (Cathepsin L) , E: CD147 , and F: GRP78 across lung epithelial cell populations in healthy subjects (see methods for dataset reference for cell population markers).

    Article Snippet: Collectively, the profiling of antibodies by immunoblot of airway epithelial cells revealed distinct band patterns demonstrative of antibody specificity for ACE2, CD147, and GRP78, and to a lesser extent for TMPRSS2.

    Techniques: Expressing

    Immunoblot analysis of ACE2, TMPRSS2, CD147, and GRP78 protein expression in human airway epithelial cell protein lysates. A: ACE2 with single band for predicted molecular weight of 110kDa. B: TMPRSS2 with multiple bands including a dominant band at predicted molecular weight of 57 kDa. C: CD147 with a single broad band around predicted molecular weight of 55kDa. D: GRP78 with a single band at predicted molecular weight of 78 kDa. Lanes 1-3: Calu-3 cells. Lanes 4-6: Primary human airway epithelial cells. Lanes 7-9: HBEC-6KT. All cells grown under submerged monolayer conditions, with n=3 independent passages (Calu-3 or HBEC-6KT) or donor samples (Primary human airway epithelial cells – non-smoker, healthy subjects). For each independent blot of each protein, all of the same samples were run. Total protein loading control provided to demonstrate equal protein loaded for each sample.

    Journal: bioRxiv

    Article Title: Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue

    doi: 10.1101/2020.04.07.030742

    Figure Lengend Snippet: Immunoblot analysis of ACE2, TMPRSS2, CD147, and GRP78 protein expression in human airway epithelial cell protein lysates. A: ACE2 with single band for predicted molecular weight of 110kDa. B: TMPRSS2 with multiple bands including a dominant band at predicted molecular weight of 57 kDa. C: CD147 with a single broad band around predicted molecular weight of 55kDa. D: GRP78 with a single band at predicted molecular weight of 78 kDa. Lanes 1-3: Calu-3 cells. Lanes 4-6: Primary human airway epithelial cells. Lanes 7-9: HBEC-6KT. All cells grown under submerged monolayer conditions, with n=3 independent passages (Calu-3 or HBEC-6KT) or donor samples (Primary human airway epithelial cells – non-smoker, healthy subjects). For each independent blot of each protein, all of the same samples were run. Total protein loading control provided to demonstrate equal protein loaded for each sample.

    Article Snippet: Collectively, the profiling of antibodies by immunoblot of airway epithelial cells revealed distinct band patterns demonstrative of antibody specificity for ACE2, CD147, and GRP78, and to a lesser extent for TMPRSS2.

    Techniques: Expressing, Molecular Weight

    Microarray expression profiles of candidate SARS-CoV-2 receptor genes in upper and lower airways. A: Normalized log 2 expression levels for ACE2, TMPRSS2, ADAM17, CTSL, CD147 , and GRP78 genes compared across the upper airway (nasal, grey) and lower airways (trachea, green; large airway, blue; small airway, orange). CDH1 gene expression level is included as a positive control. B: Statistical values for comparisons for each gene at each airway generation. *= p

    Journal: bioRxiv

    Article Title: Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue

    doi: 10.1101/2020.04.07.030742

    Figure Lengend Snippet: Microarray expression profiles of candidate SARS-CoV-2 receptor genes in upper and lower airways. A: Normalized log 2 expression levels for ACE2, TMPRSS2, ADAM17, CTSL, CD147 , and GRP78 genes compared across the upper airway (nasal, grey) and lower airways (trachea, green; large airway, blue; small airway, orange). CDH1 gene expression level is included as a positive control. B: Statistical values for comparisons for each gene at each airway generation. *= p

    Article Snippet: Collectively, the profiling of antibodies by immunoblot of airway epithelial cells revealed distinct band patterns demonstrative of antibody specificity for ACE2, CD147, and GRP78, and to a lesser extent for TMPRSS2.

    Techniques: Microarray, Expressing, Positive Control

    Microarray expression profiles of candidate SARS-CoV-2 receptor genes in lower airway epithelial cells analyzed by age and sex. A: Clustered heatmap of log 2 expression levels from 504 GEO samples, annotated by age, sex, and microarray chip platform. Expression values reflect signal intensities, indicating lowest detected expression of ACE2 and highest expression of GRP78 and CDH1 . B: Per-gene boxplots of expression levels separated by sex. C: Plots of gene expression levels versus age, with linear regression lines of best fit. A weak negative correlation ( r = −0.20, p = 0.015) was detected for ACE2 in the second dataset. Correlations were performed separately between platforms because of differences in their age distributions.

    Journal: bioRxiv

    Article Title: Gene expression and in situ protein profiling of candidate SARS-CoV-2 receptors in human airway epithelial cells and lung tissue

    doi: 10.1101/2020.04.07.030742

    Figure Lengend Snippet: Microarray expression profiles of candidate SARS-CoV-2 receptor genes in lower airway epithelial cells analyzed by age and sex. A: Clustered heatmap of log 2 expression levels from 504 GEO samples, annotated by age, sex, and microarray chip platform. Expression values reflect signal intensities, indicating lowest detected expression of ACE2 and highest expression of GRP78 and CDH1 . B: Per-gene boxplots of expression levels separated by sex. C: Plots of gene expression levels versus age, with linear regression lines of best fit. A weak negative correlation ( r = −0.20, p = 0.015) was detected for ACE2 in the second dataset. Correlations were performed separately between platforms because of differences in their age distributions.

    Article Snippet: Collectively, the profiling of antibodies by immunoblot of airway epithelial cells revealed distinct band patterns demonstrative of antibody specificity for ACE2, CD147, and GRP78, and to a lesser extent for TMPRSS2.

    Techniques: Microarray, Expressing, Chromatin Immunoprecipitation

    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

    Estradiol treatment upregulates the expression of endoplasmic reticulum stress (ERS) related proteins in esophageal cancer cell line EC109. The cells were cultured in serum-free Roswell Park Memorial Institute (RPMI) 1640 medium for 24 h, and treated with 17-estradiol (E2), E2 and ICI 182,780 (E2 + ICI), E2 and 4-phenylbutyric acid (4-PBA), and tunicamycin (TM) for 24 h, respectively. (A) GRP78 proteins in EC109 cells under an Olympus laser confocal microscope. (B–E) The expressions of GRP78, ATF6, IRE1α, and PERK in EC109 cells after various treatments. Densitometric values were normalized to β-actin. The results are expressed as the means ± SEM, n = 3. * P

    Journal: Frontiers in Endocrinology

    Article Title: Interaction of Estradiol and Endoplasmic Reticulum Stress in the Development of Esophageal Carcinoma

    doi: 10.3389/fendo.2020.00410

    Figure Lengend Snippet: Estradiol treatment upregulates the expression of endoplasmic reticulum stress (ERS) related proteins in esophageal cancer cell line EC109. The cells were cultured in serum-free Roswell Park Memorial Institute (RPMI) 1640 medium for 24 h, and treated with 17-estradiol (E2), E2 and ICI 182,780 (E2 + ICI), E2 and 4-phenylbutyric acid (4-PBA), and tunicamycin (TM) for 24 h, respectively. (A) GRP78 proteins in EC109 cells under an Olympus laser confocal microscope. (B–E) The expressions of GRP78, ATF6, IRE1α, and PERK in EC109 cells after various treatments. Densitometric values were normalized to β-actin. The results are expressed as the means ± SEM, n = 3. * P

    Article Snippet: After permeabilization with Triton X-100, cells were blocked with 5% normal bovine serum albumin for 30 min and incubated with antiglucose regulated protein 78 (GRP78) (1:500, ab21685, Abcam), antiestrogen receptor α (ERα) (1:100, ab32063, Abcam), and anti-ERβ (1:100, ab212351, Abcam), respectively, at 4°C overnight.

    Techniques: Expressing, Cell Culture, Microscopy

    Estradiol treatment inhibits the viability and migration of esophageal cancer cells through the activation of endoplasmic reticulum stress (ERS). ICI, an inhibitor of estrogen nuclear receptors; 4-PBA, an inhibitor of ERS; tunicamycin (TM), an agonist of ERS; GRP78, ATF6, PERK, and IRE1 were ERS-related proteins.

    Journal: Frontiers in Endocrinology

    Article Title: Interaction of Estradiol and Endoplasmic Reticulum Stress in the Development of Esophageal Carcinoma

    doi: 10.3389/fendo.2020.00410

    Figure Lengend Snippet: Estradiol treatment inhibits the viability and migration of esophageal cancer cells through the activation of endoplasmic reticulum stress (ERS). ICI, an inhibitor of estrogen nuclear receptors; 4-PBA, an inhibitor of ERS; tunicamycin (TM), an agonist of ERS; GRP78, ATF6, PERK, and IRE1 were ERS-related proteins.

    Article Snippet: After permeabilization with Triton X-100, cells were blocked with 5% normal bovine serum albumin for 30 min and incubated with antiglucose regulated protein 78 (GRP78) (1:500, ab21685, Abcam), antiestrogen receptor α (ERα) (1:100, ab32063, Abcam), and anti-ERβ (1:100, ab212351, Abcam), respectively, at 4°C overnight.

    Techniques: Migration, Activation Assay

    LRRK2 supports GRP78-mediated cell survival. ( A ) Quantification of the populations of cells with sub-G0 DNA content in cultures of SH-SY5Y cells transfected with a control vector (Control) and MIX LRRK2 KD SH-SY5Y cells (LRRK2 KD) treated with 0, 3 or 6 µM tunicamycin for 16 hours. Data represent the mean ± SEM of 3 independent experiments. ***, p

    Journal: PLoS ONE

    Article Title: Dysregulated LRRK2 Signaling in Response to Endoplasmic Reticulum Stress Leads to Dopaminergic Neuron Degeneration in C. elegans

    doi: 10.1371/journal.pone.0022354

    Figure Lengend Snippet: LRRK2 supports GRP78-mediated cell survival. ( A ) Quantification of the populations of cells with sub-G0 DNA content in cultures of SH-SY5Y cells transfected with a control vector (Control) and MIX LRRK2 KD SH-SY5Y cells (LRRK2 KD) treated with 0, 3 or 6 µM tunicamycin for 16 hours. Data represent the mean ± SEM of 3 independent experiments. ***, p

    Article Snippet: ; anti-p38, phospho-specific anti-P-p38, anti-GRP78, anti-eIF2α, phospho-specific anti-P-eIF2α, and anti-MKK6 (Cell Signaling, Danvers, MA); and anti-LRRK2 (Sigma).

    Techniques: Transfection, Plasmid Preparation

    LRRK2 signaling modulates GRP78 levels to potentiate cell survival after 6-OHDA exposure. ( A ) LRRK2 is involved in 6-OHDA-mediated upregulation of GRP78 protein levels. Western blot analyses were performed on lysates from SH-SY5Y cells transfected with a control vector (Control, circles) and MIX LRRK2 KD SH-SY5Y cells (LRRK2 KD, diamonds) treated with 100 µM 6-OHDA for the indicated periods of time. Representative Western blots are shown, as well as quantization of results from 3 independent experiments. Data represent the mean ± SEM of 3 independent experiments. **, p

    Journal: PLoS ONE

    Article Title: Dysregulated LRRK2 Signaling in Response to Endoplasmic Reticulum Stress Leads to Dopaminergic Neuron Degeneration in C. elegans

    doi: 10.1371/journal.pone.0022354

    Figure Lengend Snippet: LRRK2 signaling modulates GRP78 levels to potentiate cell survival after 6-OHDA exposure. ( A ) LRRK2 is involved in 6-OHDA-mediated upregulation of GRP78 protein levels. Western blot analyses were performed on lysates from SH-SY5Y cells transfected with a control vector (Control, circles) and MIX LRRK2 KD SH-SY5Y cells (LRRK2 KD, diamonds) treated with 100 µM 6-OHDA for the indicated periods of time. Representative Western blots are shown, as well as quantization of results from 3 independent experiments. Data represent the mean ± SEM of 3 independent experiments. **, p

    Article Snippet: ; anti-p38, phospho-specific anti-P-p38, anti-GRP78, anti-eIF2α, phospho-specific anti-P-eIF2α, and anti-MKK6 (Cell Signaling, Danvers, MA); and anti-LRRK2 (Sigma).

    Techniques: Western Blot, Transfection, Plasmid Preparation

    Effects of LM-1685, a non-coxib COX-2 inhibitor, on the viability and induction of ER stress-related molecules in NTUB1 and T24 cells. (A) Cells were treated with LM-1685 (0–160 µM) for 24 h. Cell viability was analyzed by MTT assay. Data are presented as means ± SD of three independents experiments. (B) Cells were treated with LM-1685 (80 and 160 µM) or celecoxib (100 µM) for 24 h. The cell lysates were harvested at each time point and analyzed by Western blotting with specific antibodies to detect ER stress-related molecules IRE-1α, CHOP, calnexin, and GRP78, and caspase-4. CF is the abbreviation of cleaved form. (C). Cells were transfected with GRP78 siRNA (10 nM) or scramble siRNA (10 nM) (as a control), and then treated with 160 µM LM-1685. The combinative effect of LM-1685 and GRP78 knockdown on apoptosis was determined by flow cytometry (FACS) with annexin V-FITC and PI labeling. The quantitative analysis of total apoptosis (early and late) population was presented. Data are presented as means ± SD of three independents experiments. * p

    Journal: PLoS ONE

    Article Title: Down-Regulation of Glucose-Regulated Protein (GRP) 78 Potentiates Cytotoxic Effect of Celecoxib in Human Urothelial Carcinoma Cells

    doi: 10.1371/journal.pone.0033615

    Figure Lengend Snippet: Effects of LM-1685, a non-coxib COX-2 inhibitor, on the viability and induction of ER stress-related molecules in NTUB1 and T24 cells. (A) Cells were treated with LM-1685 (0–160 µM) for 24 h. Cell viability was analyzed by MTT assay. Data are presented as means ± SD of three independents experiments. (B) Cells were treated with LM-1685 (80 and 160 µM) or celecoxib (100 µM) for 24 h. The cell lysates were harvested at each time point and analyzed by Western blotting with specific antibodies to detect ER stress-related molecules IRE-1α, CHOP, calnexin, and GRP78, and caspase-4. CF is the abbreviation of cleaved form. (C). Cells were transfected with GRP78 siRNA (10 nM) or scramble siRNA (10 nM) (as a control), and then treated with 160 µM LM-1685. The combinative effect of LM-1685 and GRP78 knockdown on apoptosis was determined by flow cytometry (FACS) with annexin V-FITC and PI labeling. The quantitative analysis of total apoptosis (early and late) population was presented. Data are presented as means ± SD of three independents experiments. * p

    Article Snippet: The combinative treatment of EGCG induced down-regulation of GRP78 and enhanced the celecoxib-induced cytotoxicity in NTUB1 and T24 cells ( ).

    Techniques: MTT Assay, Western Blot, Transfection, Flow Cytometry, Cytometry, FACS, Labeling

    The combinative effects of celecoxib and GRP78 knockdown on apoptosis of NTUB1 and T24 cells. (A) Cells were transfected with GRP78 siRNA (10 nM) or scramble siRNA (10 nM) (as a control); then treated with 100 µM celecoxib. The combinative effect of celecoxib and GRP78 knockdown on apoptosis was determined by flow cytometry (FACS) with annexin V-FITC and PI labeling in (a). (b) Quantitative analysis of total apoptosis (early and late) population was presented. Data are presented as means ± SD of three independents experiments. * p

    Journal: PLoS ONE

    Article Title: Down-Regulation of Glucose-Regulated Protein (GRP) 78 Potentiates Cytotoxic Effect of Celecoxib in Human Urothelial Carcinoma Cells

    doi: 10.1371/journal.pone.0033615

    Figure Lengend Snippet: The combinative effects of celecoxib and GRP78 knockdown on apoptosis of NTUB1 and T24 cells. (A) Cells were transfected with GRP78 siRNA (10 nM) or scramble siRNA (10 nM) (as a control); then treated with 100 µM celecoxib. The combinative effect of celecoxib and GRP78 knockdown on apoptosis was determined by flow cytometry (FACS) with annexin V-FITC and PI labeling in (a). (b) Quantitative analysis of total apoptosis (early and late) population was presented. Data are presented as means ± SD of three independents experiments. * p

    Article Snippet: The combinative treatment of EGCG induced down-regulation of GRP78 and enhanced the celecoxib-induced cytotoxicity in NTUB1 and T24 cells ( ).

    Techniques: Transfection, Flow Cytometry, Cytometry, FACS, Labeling

    Effects of celecoxib on the ER stress-related signaling molecules IRE-1α, GRP78, CHOP, and caspase-4 in NTUB1 and T24 cells. In (A) and (B), NTUB1 cells were exposed to mock (untreated) and 100 µM celecoxib. The cell lysates were harvested at each time point and analyzed by Western blotting with specific antibodies to detect ER stress-related molecules IRE-1α, CHOP, calnexin, GRP78, and caspase-4,. CF is the abbreviation of cleaved form. (C) and (D) showed the similar experiments performed in T24 cells. Results shown are representative of at least four independent experiments.

    Journal: PLoS ONE

    Article Title: Down-Regulation of Glucose-Regulated Protein (GRP) 78 Potentiates Cytotoxic Effect of Celecoxib in Human Urothelial Carcinoma Cells

    doi: 10.1371/journal.pone.0033615

    Figure Lengend Snippet: Effects of celecoxib on the ER stress-related signaling molecules IRE-1α, GRP78, CHOP, and caspase-4 in NTUB1 and T24 cells. In (A) and (B), NTUB1 cells were exposed to mock (untreated) and 100 µM celecoxib. The cell lysates were harvested at each time point and analyzed by Western blotting with specific antibodies to detect ER stress-related molecules IRE-1α, CHOP, calnexin, GRP78, and caspase-4,. CF is the abbreviation of cleaved form. (C) and (D) showed the similar experiments performed in T24 cells. Results shown are representative of at least four independent experiments.

    Article Snippet: The combinative treatment of EGCG induced down-regulation of GRP78 and enhanced the celecoxib-induced cytotoxicity in NTUB1 and T24 cells ( ).

    Techniques: Western Blot

    The combinative effect of celecoxib and MG132 on cell growth and apoptosis in NTUB1 and T24 cells. Cells were incubated in the presence of 100 µM celecoxib and MG132 (0.5 and 1 µM) individually or in combination. (A) Cell viability was measured by MTT assay. (B) Cells were stained with annexin V-FITC and PI for apoptosis analysis by FACS flow cytometry. (C) Quantitative analysis of total apoptosis (early and late) population was presented. (D) and (E) Cells were incubated in the presence of 100 µM celecoxib and MG132 (1 µM) individually or in combination. Cell lysates were harvested and analyzed by Western blotting with specific antibodies against, caspase-3, 7, 8, 9, PARP, CHOP, GRP78, and ubiquitin. In A and C, data are presented as means ± SD of three independents experiments. * p

    Journal: PLoS ONE

    Article Title: Down-Regulation of Glucose-Regulated Protein (GRP) 78 Potentiates Cytotoxic Effect of Celecoxib in Human Urothelial Carcinoma Cells

    doi: 10.1371/journal.pone.0033615

    Figure Lengend Snippet: The combinative effect of celecoxib and MG132 on cell growth and apoptosis in NTUB1 and T24 cells. Cells were incubated in the presence of 100 µM celecoxib and MG132 (0.5 and 1 µM) individually or in combination. (A) Cell viability was measured by MTT assay. (B) Cells were stained with annexin V-FITC and PI for apoptosis analysis by FACS flow cytometry. (C) Quantitative analysis of total apoptosis (early and late) population was presented. (D) and (E) Cells were incubated in the presence of 100 µM celecoxib and MG132 (1 µM) individually or in combination. Cell lysates were harvested and analyzed by Western blotting with specific antibodies against, caspase-3, 7, 8, 9, PARP, CHOP, GRP78, and ubiquitin. In A and C, data are presented as means ± SD of three independents experiments. * p

    Article Snippet: The combinative treatment of EGCG induced down-regulation of GRP78 and enhanced the celecoxib-induced cytotoxicity in NTUB1 and T24 cells ( ).

    Techniques: Incubation, MTT Assay, Staining, FACS, Flow Cytometry, Cytometry, Western Blot

    Effects of darbepoetin alfa on left ventricular myocardial GRP78, CHOP, ATF6, and caspase-12 in the Sham– and β 1 -EC II –immunized rabbits. Representative immunoblots are shown on the left to the bar graphs. N=6 animals in each group. Bar denote SEM. *P

    Journal: Journal of molecular and cellular cardiology

    Article Title: Darbepoetin Alfa Exerts A Cardioprotective Effect in Autoimmune Cardiomyopathy via Reduction of ER Stress and Activation of the PI3K/Akt and STAT3 Pathways

    doi: 10.1016/j.yjmcc.2008.05.010

    Figure Lengend Snippet: Effects of darbepoetin alfa on left ventricular myocardial GRP78, CHOP, ATF6, and caspase-12 in the Sham– and β 1 -EC II –immunized rabbits. Representative immunoblots are shown on the left to the bar graphs. N=6 animals in each group. Bar denote SEM. *P

    Article Snippet: Blots was then probed with the following antibodies: Monoclonal anti-caspase-12 (1:500, Sigma-Aldrich), anti-GRP78 (1:2000, BD Biosciences, San Jose, CA), anti-CHOP (1: 200, Santa Cruz), anti-phospho-Akt (Ser472), anti-Akt (1: 1000, Cell Signaling), anti-phospho-STAT3 (Ser727), anti-STAT3 (1:500 each, Cell Signaling), anti-Bcl2 (1: 1000, Santa Cruz), polyclonal anti-Bax (1:1000, Cell Signaling), anti-phospho-P38 MAPK (1:500, Cell Signaling), anti-p38 MAPK (1:1000, Cell Signaling), anti-phospho-ERK1/2 and anti-ERK1/2 (both 1:1000, Cell Signaling), and anti-EpoR antibody (1:1000, Santa Cruz).

    Techniques: Western Blot

    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

    HCV E2 inhibited the interaction between grp78 and AIMP1/p43. A. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody. B. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells along with 2 or 4 µg of plasmid expressing HCV E2 and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody. C. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells along with plasmid expressing HCV E2, E1, core, or NS5A and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody.

    Journal: PLoS ONE

    Article Title: Degradation of AIMP1/p43 Induced by Hepatitis C Virus E2 Leads to Upregulation of TGF-? Signaling and Increase in Surface Expression of gp96

    doi: 10.1371/journal.pone.0096302

    Figure Lengend Snippet: HCV E2 inhibited the interaction between grp78 and AIMP1/p43. A. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody. B. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells along with 2 or 4 µg of plasmid expressing HCV E2 and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody. C. Plasmid expressing V5-tagged grp78 and plasmid expressing AIMP1/p43 were transfected to HEK 293 cells along with plasmid expressing HCV E2, E1, core, or NS5A and grown. Cell lysate was subjected to immunoprecipitation using anti-AIMP1/p43 antibody.

    Article Snippet: HCV E2 inhibited interaction between grp78 and AIMP1/p43 Above experiments showed that grp78 acted as a stabilizer for AIMP1/p43.

    Techniques: Plasmid Preparation, Expressing, Transfection, Immunoprecipitation

    HCV E2 inhibited stabilization of AIMP1/p43 by grp78. A. Plasmid expressing grp78 was transfected to Huh-7 cells and grown. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody. β-tubulin was used as a loading control. EV; empty vector. B. grp78 siRNA or negative control siRNA was transfected to Huh-7 cells and grown. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody. C. grp7 siRNA was transfected to Huh-7 cells at indicated amounts. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody.

    Journal: PLoS ONE

    Article Title: Degradation of AIMP1/p43 Induced by Hepatitis C Virus E2 Leads to Upregulation of TGF-? Signaling and Increase in Surface Expression of gp96

    doi: 10.1371/journal.pone.0096302

    Figure Lengend Snippet: HCV E2 inhibited stabilization of AIMP1/p43 by grp78. A. Plasmid expressing grp78 was transfected to Huh-7 cells and grown. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody. β-tubulin was used as a loading control. EV; empty vector. B. grp78 siRNA or negative control siRNA was transfected to Huh-7 cells and grown. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody. C. grp7 siRNA was transfected to Huh-7 cells at indicated amounts. Cell lysate was subjected to western blot using anti AIMP1/p43 antibody and anti-grp78 antibody.

    Article Snippet: HCV E2 inhibited interaction between grp78 and AIMP1/p43 Above experiments showed that grp78 acted as a stabilizer for AIMP1/p43.

    Techniques: Plasmid Preparation, Expressing, Transfection, Western Blot, Negative Control

    Semi-quantitative analysis of immunohistochemical staining of GRP78, p-ERK and caspase-12 in the kidney, based on the percentage of positively stained areas in the tubules and the intensity of staining. Animals were divided into four groups: the control group (N), the cisplatin (CP) group (C), the grape seed proanthocyanidin extract (GSPE) group (G) and the CP+GSPE group (C+G). The expression of GRP78, p-ERK and caspase-12 was significantly increased in the C group compared to the N group and significantly decreased in the C+G group compared to the C group. * Statistically significant difference compared to the N group (p

    Journal: Molecular Medicine Reports

    Article Title: Grape seed proanthocyanidin extract protects from cisplatin-induced nephrotoxicity by inhibiting endoplasmic reticulum stress-induced apoptosis

    doi: 10.3892/mmr.2014.1883

    Figure Lengend Snippet: Semi-quantitative analysis of immunohistochemical staining of GRP78, p-ERK and caspase-12 in the kidney, based on the percentage of positively stained areas in the tubules and the intensity of staining. Animals were divided into four groups: the control group (N), the cisplatin (CP) group (C), the grape seed proanthocyanidin extract (GSPE) group (G) and the CP+GSPE group (C+G). The expression of GRP78, p-ERK and caspase-12 was significantly increased in the C group compared to the N group and significantly decreased in the C+G group compared to the C group. * Statistically significant difference compared to the N group (p

    Article Snippet: Primary antibodies used in this study were: rabbit anti-GRP78, rabbit anti-caspase-12 (Abcam Ltd., Hong Kong, China), rabbit anti-p-ERK, rabbit anti-ERK (Cell Signaling Technology, Inc., Danvers, MA, USA) and mouse anti-β-actin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA).

    Techniques: Immunohistochemistry, Staining, Expressing

    Western blot analysis for GRP78, p-ERK and caspase-12, and quantification of corresponding protein levels. Animals were divided into four groups: the control group (N), the cisplatin (CP) group (C), the grape seed proanthocyanidin extract (GSPE) group (G) and the CP+GSPE group (C+G). Data are expressed as mean ± SD levels relative to β-actin. * Statistically significant difference compared to the N group (P

    Journal: Molecular Medicine Reports

    Article Title: Grape seed proanthocyanidin extract protects from cisplatin-induced nephrotoxicity by inhibiting endoplasmic reticulum stress-induced apoptosis

    doi: 10.3892/mmr.2014.1883

    Figure Lengend Snippet: Western blot analysis for GRP78, p-ERK and caspase-12, and quantification of corresponding protein levels. Animals were divided into four groups: the control group (N), the cisplatin (CP) group (C), the grape seed proanthocyanidin extract (GSPE) group (G) and the CP+GSPE group (C+G). Data are expressed as mean ± SD levels relative to β-actin. * Statistically significant difference compared to the N group (P

    Article Snippet: Primary antibodies used in this study were: rabbit anti-GRP78, rabbit anti-caspase-12 (Abcam Ltd., Hong Kong, China), rabbit anti-p-ERK, rabbit anti-ERK (Cell Signaling Technology, Inc., Danvers, MA, USA) and mouse anti-β-actin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA).

    Techniques: Western Blot

    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

    Synaptic markers in VD. Pre-synaptic, pos-synaptic and mitochondrial proteins were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of VD patients and controls. The levels of synaptophysin, PSD95 and HSP60 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, *p

    Journal: Scientific Reports

    Article Title: Differential protein expression in diverse brain areas of Parkinson’s and Alzheimer’s disease patients

    doi: 10.1038/s41598-020-70174-z

    Figure Lengend Snippet: Synaptic markers in VD. Pre-synaptic, pos-synaptic and mitochondrial proteins were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of VD patients and controls. The levels of synaptophysin, PSD95 and HSP60 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, *p

    Article Snippet: The primary antibodies used were the following: 1:1,000 polyclonal anti-alpha-SNCA, oligomer specific Syn-33 from Sigma (St. Louis, MO, USA) (cat number: ABN2265); 1:1,000 polyclonal anti-LC3B (microtubule-associated protein 1A/1B-light chain 3) from Cell Signaling (Danvers, MA, USA) (cat number: 3868); 1:16,000 monoclonal anti-acetylated α-tubulin from Sigma (St. Louis, MO, USA) (cat number: T7451); 1:1,000 monoclonal anti-synaptophysin from Sigma (St. Louis, MO, USA) (cat number: S5768); 1:1,000 anti-PSD95 (postsynaptic density protein 95) antibody from Abcam (Cambridge; UK) (cat number: ab18258); 1:1,000 anti-Lamp2A (lysosomal-associated membrane protein 2A) from Novus Biologicals (Littleton, CO, USA) (cat number: ab37024); 1:750 anti-Tau Acetyl K280 from AnaSpec (Fremont, CA, USA) (cat number: AS-56077); 1:1,000 anti-Beclin1 from Cell Signaling (Danvers, MA, USA) (cat number: 3738); 1:1,000 anti-p62 from Sigma (St. Louis, MO, USA) (cat number: P0067); 1:1,000 anti-Lamp1 (lysosomal-associated membrane protein 1) from Cell Signaling (Danvers, MA, USA) (cat number: 3243); 1:750 anti-phospho-Tau Ser396 from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-101815); 1:1,000 anti-Aβ from Cell Signaling (Danvers, MA, USA) (cat number: 8243); 1:1,000 anti-cathepsin D (CatD) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-377124); 1:1,000 anti-Hsp60 from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 611563); 1:1,000 monoclonal anti-Tau, clone Tau46 from Sigma (St. Louis, MO, USA) (cat number: T9450); 1:1,000 anti-BiP/GRP78 (78-kDa Glucose-regulated protein) from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 610979); 1:1,000 anti-ATF4 (Activating transcription factor 4) from Cell Signaling (Danvers, MA, USA) (cat number: 97038).

    Techniques:

    ER Stress in PD. UPR markers were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of sporadic PD patients and controls. The levels of GRP78 and ATF4 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ). Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, **p

    Journal: Scientific Reports

    Article Title: Differential protein expression in diverse brain areas of Parkinson’s and Alzheimer’s disease patients

    doi: 10.1038/s41598-020-70174-z

    Figure Lengend Snippet: ER Stress in PD. UPR markers were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of sporadic PD patients and controls. The levels of GRP78 and ATF4 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ). Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, **p

    Article Snippet: The primary antibodies used were the following: 1:1,000 polyclonal anti-alpha-SNCA, oligomer specific Syn-33 from Sigma (St. Louis, MO, USA) (cat number: ABN2265); 1:1,000 polyclonal anti-LC3B (microtubule-associated protein 1A/1B-light chain 3) from Cell Signaling (Danvers, MA, USA) (cat number: 3868); 1:16,000 monoclonal anti-acetylated α-tubulin from Sigma (St. Louis, MO, USA) (cat number: T7451); 1:1,000 monoclonal anti-synaptophysin from Sigma (St. Louis, MO, USA) (cat number: S5768); 1:1,000 anti-PSD95 (postsynaptic density protein 95) antibody from Abcam (Cambridge; UK) (cat number: ab18258); 1:1,000 anti-Lamp2A (lysosomal-associated membrane protein 2A) from Novus Biologicals (Littleton, CO, USA) (cat number: ab37024); 1:750 anti-Tau Acetyl K280 from AnaSpec (Fremont, CA, USA) (cat number: AS-56077); 1:1,000 anti-Beclin1 from Cell Signaling (Danvers, MA, USA) (cat number: 3738); 1:1,000 anti-p62 from Sigma (St. Louis, MO, USA) (cat number: P0067); 1:1,000 anti-Lamp1 (lysosomal-associated membrane protein 1) from Cell Signaling (Danvers, MA, USA) (cat number: 3243); 1:750 anti-phospho-Tau Ser396 from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-101815); 1:1,000 anti-Aβ from Cell Signaling (Danvers, MA, USA) (cat number: 8243); 1:1,000 anti-cathepsin D (CatD) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-377124); 1:1,000 anti-Hsp60 from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 611563); 1:1,000 monoclonal anti-Tau, clone Tau46 from Sigma (St. Louis, MO, USA) (cat number: T9450); 1:1,000 anti-BiP/GRP78 (78-kDa Glucose-regulated protein) from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 610979); 1:1,000 anti-ATF4 (Activating transcription factor 4) from Cell Signaling (Danvers, MA, USA) (cat number: 97038).

    Techniques:

    Synaptic markers in AD. Pre-synaptic, pos-synaptic and mitochondrial proteins were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of sporadic AD patients and controls. The levels of synaptophysin, PSD95 and HSP60 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5). Full length blots are presented in the Supplementary Information.

    Journal: Scientific Reports

    Article Title: Differential protein expression in diverse brain areas of Parkinson’s and Alzheimer’s disease patients

    doi: 10.1038/s41598-020-70174-z

    Figure Lengend Snippet: Synaptic markers in AD. Pre-synaptic, pos-synaptic and mitochondrial proteins were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of sporadic AD patients and controls. The levels of synaptophysin, PSD95 and HSP60 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5). Full length blots are presented in the Supplementary Information.

    Article Snippet: The primary antibodies used were the following: 1:1,000 polyclonal anti-alpha-SNCA, oligomer specific Syn-33 from Sigma (St. Louis, MO, USA) (cat number: ABN2265); 1:1,000 polyclonal anti-LC3B (microtubule-associated protein 1A/1B-light chain 3) from Cell Signaling (Danvers, MA, USA) (cat number: 3868); 1:16,000 monoclonal anti-acetylated α-tubulin from Sigma (St. Louis, MO, USA) (cat number: T7451); 1:1,000 monoclonal anti-synaptophysin from Sigma (St. Louis, MO, USA) (cat number: S5768); 1:1,000 anti-PSD95 (postsynaptic density protein 95) antibody from Abcam (Cambridge; UK) (cat number: ab18258); 1:1,000 anti-Lamp2A (lysosomal-associated membrane protein 2A) from Novus Biologicals (Littleton, CO, USA) (cat number: ab37024); 1:750 anti-Tau Acetyl K280 from AnaSpec (Fremont, CA, USA) (cat number: AS-56077); 1:1,000 anti-Beclin1 from Cell Signaling (Danvers, MA, USA) (cat number: 3738); 1:1,000 anti-p62 from Sigma (St. Louis, MO, USA) (cat number: P0067); 1:1,000 anti-Lamp1 (lysosomal-associated membrane protein 1) from Cell Signaling (Danvers, MA, USA) (cat number: 3243); 1:750 anti-phospho-Tau Ser396 from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-101815); 1:1,000 anti-Aβ from Cell Signaling (Danvers, MA, USA) (cat number: 8243); 1:1,000 anti-cathepsin D (CatD) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-377124); 1:1,000 anti-Hsp60 from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 611563); 1:1,000 monoclonal anti-Tau, clone Tau46 from Sigma (St. Louis, MO, USA) (cat number: T9450); 1:1,000 anti-BiP/GRP78 (78-kDa Glucose-regulated protein) from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 610979); 1:1,000 anti-ATF4 (Activating transcription factor 4) from Cell Signaling (Danvers, MA, USA) (cat number: 97038).

    Techniques:

    ER Stress in AD. UPR markers were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of sporadic AD patients and controls. The levels of GRP78 and ATF4 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5). Full length blots are presented in the Supplementary Information.

    Journal: Scientific Reports

    Article Title: Differential protein expression in diverse brain areas of Parkinson’s and Alzheimer’s disease patients

    doi: 10.1038/s41598-020-70174-z

    Figure Lengend Snippet: ER Stress in AD. UPR markers were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of sporadic AD patients and controls. The levels of GRP78 and ATF4 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5). Full length blots are presented in the Supplementary Information.

    Article Snippet: The primary antibodies used were the following: 1:1,000 polyclonal anti-alpha-SNCA, oligomer specific Syn-33 from Sigma (St. Louis, MO, USA) (cat number: ABN2265); 1:1,000 polyclonal anti-LC3B (microtubule-associated protein 1A/1B-light chain 3) from Cell Signaling (Danvers, MA, USA) (cat number: 3868); 1:16,000 monoclonal anti-acetylated α-tubulin from Sigma (St. Louis, MO, USA) (cat number: T7451); 1:1,000 monoclonal anti-synaptophysin from Sigma (St. Louis, MO, USA) (cat number: S5768); 1:1,000 anti-PSD95 (postsynaptic density protein 95) antibody from Abcam (Cambridge; UK) (cat number: ab18258); 1:1,000 anti-Lamp2A (lysosomal-associated membrane protein 2A) from Novus Biologicals (Littleton, CO, USA) (cat number: ab37024); 1:750 anti-Tau Acetyl K280 from AnaSpec (Fremont, CA, USA) (cat number: AS-56077); 1:1,000 anti-Beclin1 from Cell Signaling (Danvers, MA, USA) (cat number: 3738); 1:1,000 anti-p62 from Sigma (St. Louis, MO, USA) (cat number: P0067); 1:1,000 anti-Lamp1 (lysosomal-associated membrane protein 1) from Cell Signaling (Danvers, MA, USA) (cat number: 3243); 1:750 anti-phospho-Tau Ser396 from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-101815); 1:1,000 anti-Aβ from Cell Signaling (Danvers, MA, USA) (cat number: 8243); 1:1,000 anti-cathepsin D (CatD) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-377124); 1:1,000 anti-Hsp60 from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 611563); 1:1,000 monoclonal anti-Tau, clone Tau46 from Sigma (St. Louis, MO, USA) (cat number: T9450); 1:1,000 anti-BiP/GRP78 (78-kDa Glucose-regulated protein) from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 610979); 1:1,000 anti-ATF4 (Activating transcription factor 4) from Cell Signaling (Danvers, MA, USA) (cat number: 97038).

    Techniques:

    ER Stress in VD. UPR markers were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of VD patients and controls. The levels of GRP78 and ATF4 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, **p

    Journal: Scientific Reports

    Article Title: Differential protein expression in diverse brain areas of Parkinson’s and Alzheimer’s disease patients

    doi: 10.1038/s41598-020-70174-z

    Figure Lengend Snippet: ER Stress in VD. UPR markers were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of VD patients and controls. The levels of GRP78 and ATF4 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, **p

    Article Snippet: The primary antibodies used were the following: 1:1,000 polyclonal anti-alpha-SNCA, oligomer specific Syn-33 from Sigma (St. Louis, MO, USA) (cat number: ABN2265); 1:1,000 polyclonal anti-LC3B (microtubule-associated protein 1A/1B-light chain 3) from Cell Signaling (Danvers, MA, USA) (cat number: 3868); 1:16,000 monoclonal anti-acetylated α-tubulin from Sigma (St. Louis, MO, USA) (cat number: T7451); 1:1,000 monoclonal anti-synaptophysin from Sigma (St. Louis, MO, USA) (cat number: S5768); 1:1,000 anti-PSD95 (postsynaptic density protein 95) antibody from Abcam (Cambridge; UK) (cat number: ab18258); 1:1,000 anti-Lamp2A (lysosomal-associated membrane protein 2A) from Novus Biologicals (Littleton, CO, USA) (cat number: ab37024); 1:750 anti-Tau Acetyl K280 from AnaSpec (Fremont, CA, USA) (cat number: AS-56077); 1:1,000 anti-Beclin1 from Cell Signaling (Danvers, MA, USA) (cat number: 3738); 1:1,000 anti-p62 from Sigma (St. Louis, MO, USA) (cat number: P0067); 1:1,000 anti-Lamp1 (lysosomal-associated membrane protein 1) from Cell Signaling (Danvers, MA, USA) (cat number: 3243); 1:750 anti-phospho-Tau Ser396 from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-101815); 1:1,000 anti-Aβ from Cell Signaling (Danvers, MA, USA) (cat number: 8243); 1:1,000 anti-cathepsin D (CatD) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-377124); 1:1,000 anti-Hsp60 from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 611563); 1:1,000 monoclonal anti-Tau, clone Tau46 from Sigma (St. Louis, MO, USA) (cat number: T9450); 1:1,000 anti-BiP/GRP78 (78-kDa Glucose-regulated protein) from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 610979); 1:1,000 anti-ATF4 (Activating transcription factor 4) from Cell Signaling (Danvers, MA, USA) (cat number: 97038).

    Techniques:

    Synaptic markers in PD. Pre-synaptic, pos-synaptic and mitochondrial proteins were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of sporadic PD patients and controls. The levels of synaptophysin, PSD95 and HSP60 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, *p

    Journal: Scientific Reports

    Article Title: Differential protein expression in diverse brain areas of Parkinson’s and Alzheimer’s disease patients

    doi: 10.1038/s41598-020-70174-z

    Figure Lengend Snippet: Synaptic markers in PD. Pre-synaptic, pos-synaptic and mitochondrial proteins were determined in post-mortem human brain samples from SNpc, Hippocampus and Cortex of sporadic PD patients and controls. The levels of synaptophysin, PSD95 and HSP60 were determined in: ( A ) SNpc; ( B ) Hippocampus and ( C ) Cortex brain tissue homogenates. ( D ) Densitometric analysis of the levels of GRP78 and ATF4. The blots were re-probed for α-tubulin to confirm equal protein loading Values are mean ± SEM (n = 5, *p

    Article Snippet: The primary antibodies used were the following: 1:1,000 polyclonal anti-alpha-SNCA, oligomer specific Syn-33 from Sigma (St. Louis, MO, USA) (cat number: ABN2265); 1:1,000 polyclonal anti-LC3B (microtubule-associated protein 1A/1B-light chain 3) from Cell Signaling (Danvers, MA, USA) (cat number: 3868); 1:16,000 monoclonal anti-acetylated α-tubulin from Sigma (St. Louis, MO, USA) (cat number: T7451); 1:1,000 monoclonal anti-synaptophysin from Sigma (St. Louis, MO, USA) (cat number: S5768); 1:1,000 anti-PSD95 (postsynaptic density protein 95) antibody from Abcam (Cambridge; UK) (cat number: ab18258); 1:1,000 anti-Lamp2A (lysosomal-associated membrane protein 2A) from Novus Biologicals (Littleton, CO, USA) (cat number: ab37024); 1:750 anti-Tau Acetyl K280 from AnaSpec (Fremont, CA, USA) (cat number: AS-56077); 1:1,000 anti-Beclin1 from Cell Signaling (Danvers, MA, USA) (cat number: 3738); 1:1,000 anti-p62 from Sigma (St. Louis, MO, USA) (cat number: P0067); 1:1,000 anti-Lamp1 (lysosomal-associated membrane protein 1) from Cell Signaling (Danvers, MA, USA) (cat number: 3243); 1:750 anti-phospho-Tau Ser396 from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-101815); 1:1,000 anti-Aβ from Cell Signaling (Danvers, MA, USA) (cat number: 8243); 1:1,000 anti-cathepsin D (CatD) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) (cat number: sc-377124); 1:1,000 anti-Hsp60 from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 611563); 1:1,000 monoclonal anti-Tau, clone Tau46 from Sigma (St. Louis, MO, USA) (cat number: T9450); 1:1,000 anti-BiP/GRP78 (78-kDa Glucose-regulated protein) from BD Transduction Laboratories (San Jose, CA, USA) (cat number: 610979); 1:1,000 anti-ATF4 (Activating transcription factor 4) from Cell Signaling (Danvers, MA, USA) (cat number: 97038).

    Techniques: