Journal: Annals of Laboratory Medicine

Article Title: In vitro Stability of Heat Shock Protein 27 in Serum and Plasma Under Different Pre-analytical Conditions: Implications for Large-Scale Clinical Studies

doi: 10.3343/alm.2016.36.4.353

Figure Lengend Snippet: Stability results for HSP27: effects of immediate and delayed processing of blood samples with storage at 4℃

Article Snippet: Circulating HSP27 concentrations were measured by using the Human Total HSP27 DuoSet IC ELISA (Catalog No. DYC1580, R&D Systems, Minneapolis, MN, USA).

Techniques: Clinical Proteomics

Journal: Annals of Laboratory Medicine

Article Title: In vitro Stability of Heat Shock Protein 27 in Serum and Plasma Under Different Pre-analytical Conditions: Implications for Large-Scale Clinical Studies

doi: 10.3343/alm.2016.36.4.353

Figure Lengend Snippet: Stability results for HSP27: effects of repeated freeze-thaw cycles with storage of serum/plasma samples at -80℃

Article Snippet: Circulating HSP27 concentrations were measured by using the Human Total HSP27 DuoSet IC ELISA (Catalog No. DYC1580, R&D Systems, Minneapolis, MN, USA).

Techniques: Clinical Proteomics

Journal: Annals of Laboratory Medicine

Article Title: In vitro Stability of Heat Shock Protein 27 in Serum and Plasma Under Different Pre-analytical Conditions: Implications for Large-Scale Clinical Studies

doi: 10.3343/alm.2016.36.4.353

Figure Lengend Snippet: In vitro stability of HSP27 under different pre-analytical conditions: (A) relative analyte stability in serum samples and (B) plasma samples stored for 2-6 hr at 4℃ with immediate and delayed sample processing; and (C) the effect of repeated freeze-thaw cycles on HSP27 serum and plasma concentrations. Graphs show relative analyte recoveries at distinct time points (each dot represents the mean analyte concentrations relative to the baseline values of 10 healthy individuals; whiskers indicate standard deviation).

Article Snippet: Circulating HSP27 concentrations were measured by using the Human Total HSP27 DuoSet IC ELISA (Catalog No. DYC1580, R&D Systems, Minneapolis, MN, USA).

Techniques: In Vitro, Clinical Proteomics, Standard Deviation

Journal: Journal of orofacial orthopedics = Fortschritte der Kieferorthopadie : Organ/official journal Deutsche Gesellschaft fur Kieferorthopadie

Article Title: Impact of phosphorylation of heat shock protein 27 on the expression profile of periodontal ligament fibroblasts during mechanical strain.

doi: 10.1007/s00056-022-00391-w

Figure Lengend Snippet: Fig. 3 Heat shock protein 27 (HSP27) phosphorylation was upregulated after compression and transfection of pHSP27+ (a). Transfection of pHSP27+ reduced the cell number (b) and increased lactate dehydrogenase (LDH) release (c) in periodontal ligament fibroblasts (PDLF) without and with compression. Statistics: Welch analysis of variance (ANOVA) with Dunnett’s T3 multiple comparisons tests; *P< 0.05; **P< 0.01; ***P< 0.001 Abb. 3 Die HSP27(Hitzeschockprotein 27)-Phosphorylierung wurde nach Kompression und Transfektion von pHSP27+ hochreguliert (a). Die Transfektion von pHSP27+ reduzierte die Zellzahl (b) und erhöhte die LDH(Laktatdehydrogenase)-Freisetzung (c) in PDLF (parodontale Li- gamentfibroblasten) ohne und mit Kompression. Statistik: Welch-ANOVA („analysis of variance“) mit Dunnett-T3 Post-hoc-Test; *p< 0,05; **p< 0,01; ***p< 0,001

Article Snippet: This was followed by incubation with the diluted primary antibodies (HSP27 1:3000 (AF1580, R&D Systems, Minneapolis, MN, USA); pHSP27 1:500 (AF2314, R&D Systems, Minneapolis, MN, USA), PTGS2 1:2500 (PA5-1817, Thermo Fisher Scientific, Waltham, MA, USA), RANKL 1:2000 (TA306362, OriGene, Rockville, MD, USA); ACTIN 1:1000 (E1C602, ABIN274248, Antibody-online, Aachen, Germany)).

Techniques: Phospho-proteomics, Transfection

Journal: PloS one

Article Title: Human-derived physiological heat shock protein 27 complex protects brain after focal cerebral ischemia in mice.

doi: 10.1371/journal.pone.0066001

Figure Lengend Snippet: Figure 1. Characterization of hHSP27. A–B, Isolated human heat shock protein 27 (hHSP27) or recombinant HSP27 (rHSP27) proteins (1 mg or 10 mg) were separated by SDS-PAGE (A) and native-PAGE (B) and stained with Coomassie brilliant blue. C, hHSP27 proteins were immunoblotted with antibodies against HSP27, phosphorylated S15 HSP27, S78 HSP27, and S82 HSP27. D, hHSP27 proteins were separated by native-PAGE and immunoblotted with antibodies against ab-crystallin and HSP20. E, rHSP27 (10 ng), hHSP27 (10 ng), ab-crystallin (5 ng), and HSP20 (5 ng) were separated by SDS-PAGE followed by immunoblotting with antibodies against HSP27, ab-crystallin, and HSP20. doi:10.1371/journal.pone.0066001.g001

Article Snippet: The observers of the behavioral tests were blinded to the treatment groups, and mice of the various groups were randomized during a given testing period. hHSP27 and HSP27 Antibody, HSP27 Elution Peptide Administration or Recombinant HSP27 Administration Mice received intravenous injections of 50 mg of hHSP27 mixed with 50 or 500 mg of HSP27-N1 or -C1 antibody 1 h after reperfusion (n= 3 in each group), 5 or 50 mg of HSP27-N1 and - C1 peptides, which were used in the elution, intravenously 1 h after reperfusion (n= 3 in each group), or 50 mg of recombinant HSP27 (rHSP27; Acris Antibodies GmbH) 1 h after reperfusion (n= 3).

Techniques: Isolation, Recombinant, SDS Page, Clear Native PAGE, Staining, Western Blot

Journal: PloS one

Article Title: Human-derived physiological heat shock protein 27 complex protects brain after focal cerebral ischemia in mice.

doi: 10.1371/journal.pone.0066001

Figure Lengend Snippet: Figure 4. Anti-HSP27 antibody and dephosphorylation inhibit hHSP27 neuroprotective effects. A, Infarct volumes in control, hHSP27 (50 mg), hHSP27 plus HSP27 antibody cocktails, HSP27 elution peptide, recombinant HSP27, and dephosphorylated hHSP27 groups. Data are means6SEM of 3 mice in each group. **P,0.001 vs. controls. B– D, Dephosphorylated and phosphorylated hHSP27 proteins were separated by SDS- PAGE (B) and native-PAGE (C), stained with Coomassie brilliant blue (B,C), and immunoblotted with anti-phosphorylated S15 HSP27, S78 HSP27, and S82 HSP27 antibodies (D). E, Photomicrographs of infarct areas stained with cresyl violet in hHSP27 and dephosphorylated hHSP27 groups prepared 24 h after reperfusion. Scale bar = 1 mm. hHSP27, human heat shock protein. doi:10.1371/journal.pone.0066001.g004

Article Snippet: The observers of the behavioral tests were blinded to the treatment groups, and mice of the various groups were randomized during a given testing period. hHSP27 and HSP27 Antibody, HSP27 Elution Peptide Administration or Recombinant HSP27 Administration Mice received intravenous injections of 50 mg of hHSP27 mixed with 50 or 500 mg of HSP27-N1 or -C1 antibody 1 h after reperfusion (n= 3 in each group), 5 or 50 mg of HSP27-N1 and - C1 peptides, which were used in the elution, intravenously 1 h after reperfusion (n= 3 in each group), or 50 mg of recombinant HSP27 (rHSP27; Acris Antibodies GmbH) 1 h after reperfusion (n= 3).

Techniques: De-Phosphorylation Assay, Control, Recombinant, SDS Page, Clear Native PAGE, Staining

Journal: The Journal of Biological Chemistry

Article Title: Cellular stress induces non-canonical activation of the receptor tyrosine kinase EphA2 through the p38-MK2-RSK signaling pathway

doi: 10.1016/j.jbc.2023.104699

Figure Lengend Snippet: The p38-MK2-RSK-EphA2 pathway promotes U87-MG cell migration induced by TMZ. A , U87-MG cells were treated with DMSO or 100 μM temozolomide (TMZ) for 48 h. Whole-cell lysates were immunoblotted with primary antibodies against pS-EphA2, EphA2, pRSK, RSK1, RSK2, pp38, p38, pHSP27, HSP27 and β-Actin. B , U87-MG cells were transfected with siRNA against EphA2 (#1 or #2) or the negative control. After 5 h of transfection, cells were treated with DMSO or 100 μM TMZ for 72 h. Cell migration was observed using a time-lapse imaging system for 120 min. The accumulated distance of cell migration (μm) was calculated and shown in box and whisker plots. ∗ p < 0.05 by the Tukey–Kramer HSD test. Whole-cell lysates were immunoblotted with primary antibodies against EphA2 and β-Actin. C – F , U87-MG cells were treated with DMSO or 100 μM TMZ for 72 h, then treated with DMSO, 10 μM BI-D1870 ( C and E ) or 10 μM MK2 inhibitor III ( D and F ) for 2 h. Whole-cell lysates were immunoblotted with primary antibodies against pS-EphA2, EphA2, pRSK, RSK1, RSK2, pHSP27, HSP27, and β-Actin ( C and D ). Cell migration was observed using a time-lapse imaging system for 120 min and the accumulated distance of cell migration (μm) was calculated and shown in box and whisker plots ( E and F ). ∗ p < 0.05 by the Tukey–Kramer HSD test. DMSO, dimethyl sulfoxide; EphA2, ephrin type-A receptor 2; HSP27, heat shock protein 27; RSK, p90 ribosomal S6 kinase.

Article Snippet: Total antibodies against EphA2 (#6997) and phospho-specific antibodies against EphA2 (Ser-897; #6347), RSK (Ser-380; cross-reacting with RSK2 Ser-386; #11989), RSK (Thr-573; #9346), p38 (Thr-180/Tyr-182; #4511), ERK (Thr-202/Tyr-204; #9101), and MK2 (Thr-334; #3041 for and , C – E or #3007 for B and ) were purchased from Cell Signaling Technology; antibodies against total ERK (C-9), p38α (A-12), RSK1 (C-21), RSK2 (C-19), MK2 (A-11), HSP27 (F-4), α-Tubulin (B-7), and β-Actin (C-4) were from Santa Cruz Biotechnology; an antibody against FLAG (F1804) was from Merck KGaA; a phospho-antibody against HSP27 (MAB23141) was from R&D Systems.

Techniques: Migration, Transfection, Negative Control, Imaging, Whisker Assay

Journal: Scientific Reports

Article Title: HSP27 and 70 expression in thymic epithelial tumors and benign thymic alterations: diagnostic, prognostic and physiologic implications

doi: 10.1038/srep24267

Figure Lengend Snippet: HSP27 and 70 expression in TETs and analysis of clinicopathologic characteristics.

Article Snippet: HSP27 and 70 concentrations in serum samples were measured by using commercially available human HSP27 (R&D Systems, Minneapolis, MN, USA, DuoSet ® Human HSP27, DY1580) and HSP70 ELISA kits (R&D Systems, Minneapolis, MN, USA, DuoSet ® IC Total HSP70/HSPA1A, DYC1663).

Techniques: Expressing, Significance Assay

Journal: Scientific Reports

Article Title: HSP27 and 70 expression in thymic epithelial tumors and benign thymic alterations: diagnostic, prognostic and physiologic implications

doi: 10.1038/srep24267

Figure Lengend Snippet: Immunohistochemical staining of HSP27 in different histologic tumor subtypes was performed. Expression of HSP27 in thymoma WHO type A ( A ), AB ( B ), B1 ( C ), B2 ( D ), B3 ( E ), TC-SCC ( F ), MNT ( G ), and TNET ( H ), is shown (400X magnification). Hematoxylin was used for counterstaining . HSP27 , Heat Shock Protein 27; TETs , Thymic Epithelial Tumors; WHO , World Health Organization; TC , Thymic Carcinoma; SCC , Squamous Cell Carcinoma; MNT , Micronodular Thymoma; TNET , Thymic Neuroendocrine Tumor.

Article Snippet: HSP27 and 70 concentrations in serum samples were measured by using commercially available human HSP27 (R&D Systems, Minneapolis, MN, USA, DuoSet ® Human HSP27, DY1580) and HSP70 ELISA kits (R&D Systems, Minneapolis, MN, USA, DuoSet ® IC Total HSP70/HSPA1A, DYC1663).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Scientific Reports

Article Title: HSP27 and 70 expression in thymic epithelial tumors and benign thymic alterations: diagnostic, prognostic and physiologic implications

doi: 10.1038/srep24267

Figure Lengend Snippet: HSP expression in TETs and ITMIG recommended outcome measures.

Article Snippet: HSP27 and 70 concentrations in serum samples were measured by using commercially available human HSP27 (R&D Systems, Minneapolis, MN, USA, DuoSet ® Human HSP27, DY1580) and HSP70 ELISA kits (R&D Systems, Minneapolis, MN, USA, DuoSet ® IC Total HSP70/HSPA1A, DYC1663).

Techniques: Expressing

Journal: Scientific Reports

Article Title: HSP27 and 70 expression in thymic epithelial tumors and benign thymic alterations: diagnostic, prognostic and physiologic implications

doi: 10.1038/srep24267

Figure Lengend Snippet: Kaplan-Meier survival plots for HSP27 ( A ) and HSP70 ( B ) and FFR are shown. The number of patients at risk for moderate to strong and weak HSP27 and 70 expression are listed for the indicated time points. HSP27, Heat Shock Protein 27; HSP70 , Heat Shock Protein 70; TETs , Thymic Epithelial Tumors; FFR, Freedom From Recurrence.

Article Snippet: HSP27 and 70 concentrations in serum samples were measured by using commercially available human HSP27 (R&D Systems, Minneapolis, MN, USA, DuoSet ® Human HSP27, DY1580) and HSP70 ELISA kits (R&D Systems, Minneapolis, MN, USA, DuoSet ® IC Total HSP70/HSPA1A, DYC1663).

Techniques: Expressing

Journal: Scientific Reports

Article Title: HSP27 and 70 expression in thymic epithelial tumors and benign thymic alterations: diagnostic, prognostic and physiologic implications

doi: 10.1038/srep24267

Figure Lengend Snippet: Representative images show endothelial expression of HSP27 ( A ) in fetal thymus and HSP70 ( B ) in adult thymus with regular morphology (*asterisk). cTECs with intense subcapsular expression of HSP27 ( C ) in fetal thymus and HSP70 ( D ) in FTH is shown (arrows). mTECs and HCs express HSP27 ( E ) and HSP70 ( F ) in fetal thymus. Arrows indicate HCs. Staining of HSP27 ( G ) and HSP70 ( H ) in dendritic cells of GCs (GC) and TECs in MZ-like distribution (MZ) in FTH is shown. Arrows indicating HSP expressing dendritic cells in GCs (A-F, H: 400 x magnification; G: 200 x magnification). Hematoxylin was used for counterstaining. HSP27 , Heat Shock Protein 27; HSP70 , Heat Shock Protein 70; cTEC , cortical Thymic Epithelial Cells; mTEC , medullary Thymic Epithelial Cells; FTH , Follicular Thymic Hyperplasia; HC , Hassall’s Corpuscles; GC , Germinal Center; MZ , TECs with marginal zone like distribution.

Article Snippet: HSP27 and 70 concentrations in serum samples were measured by using commercially available human HSP27 (R&D Systems, Minneapolis, MN, USA, DuoSet ® Human HSP27, DY1580) and HSP70 ELISA kits (R&D Systems, Minneapolis, MN, USA, DuoSet ® IC Total HSP70/HSPA1A, DYC1663).

Techniques: Expressing, Staining

Journal: Scientific Reports

Article Title: HSP27 and 70 expression in thymic epithelial tumors and benign thymic alterations: diagnostic, prognostic and physiologic implications

doi: 10.1038/srep24267

Figure Lengend Snippet: Double-staining of Lu5-pancytokeratin (red) and HSP70 (brown) of an adult ( A ) and fetal thymus ( C ) is shown. There was complete concordance of Lu5 and HSP70 expression. Staining pattern of CD68 (PGM-1, macrophages/DCs) (red) and HSP70 (brown) double-staining is shown for an adult ( B ) and fetal ( D ) thymus. While most cells showed either distinct CD68 or HSP70 expression we found few cells that showed double staining (insert in D 630x magnification). Hematoxylin was used for counterstaining. Magnification: 400x. HSP27 , Heat Shock Protein 27; HSP70 , Heat Shock Protein 70; DCs ; dendritic cells.

Article Snippet: HSP27 and 70 concentrations in serum samples were measured by using commercially available human HSP27 (R&D Systems, Minneapolis, MN, USA, DuoSet ® Human HSP27, DY1580) and HSP70 ELISA kits (R&D Systems, Minneapolis, MN, USA, DuoSet ® IC Total HSP70/HSPA1A, DYC1663).

Techniques: Double Staining, Expressing, Staining

Journal: Scientific Reports

Article Title: HSP27 and 70 expression in thymic epithelial tumors and benign thymic alterations: diagnostic, prognostic and physiologic implications

doi: 10.1038/srep24267

Figure Lengend Snippet: Serum concentrations of HSP27 and 70 in TETs, TH and MG.

Article Snippet: HSP27 and 70 concentrations in serum samples were measured by using commercially available human HSP27 (R&D Systems, Minneapolis, MN, USA, DuoSet ® Human HSP27, DY1580) and HSP70 ELISA kits (R&D Systems, Minneapolis, MN, USA, DuoSet ® IC Total HSP70/HSPA1A, DYC1663).

Techniques:

Journal: Scientific Reports

Article Title: HSP27 and 70 expression in thymic epithelial tumors and benign thymic alterations: diagnostic, prognostic and physiologic implications

doi: 10.1038/srep24267

Figure Lengend Snippet: Serum concentrations of HSP27 ( A ) and HSP70 ( B ) in patients with TETs compared to healthy volunteers are shown. Patients with TETs were further separated into patients with TC and thymoma. Serum concentrations for HSP27 ( C ) and HSP70 ( D ) are shown. Serum levels of HSP70 in patients with TETs compared to patients with TH and volunteers is shown ( E ). Analysis of HSP70 serum concentrations in patients with TC compared to thymomas and TH is shown ( F ). Serum concentrations of HSP27 in patients with MG compared to volunteers is shown ( G ). Spearman rho correlations for HSP27 ( H ) and HSP70 ( I ) with WHO classification is shown. Pearson correlation for HSP27 and HSP70 serum concentrations are shown ( J ). Paired T-Test was performed to analyze fold decrease of HSP27 and 70 serum concentrations after tumor resection ( K ). HSP27 , Heat Shock Protein 27; HSP70 , Heat Shock Protein70; TETs , Thymic Epithelial Tumors; TH , Thymic Hyperplasia; MG , Myasthenia Gravis; TC , Thymic Carcinoma; r, correlation coefficient.

Article Snippet: HSP27 and 70 concentrations in serum samples were measured by using commercially available human HSP27 (R&D Systems, Minneapolis, MN, USA, DuoSet ® Human HSP27, DY1580) and HSP70 ELISA kits (R&D Systems, Minneapolis, MN, USA, DuoSet ® IC Total HSP70/HSPA1A, DYC1663).

Techniques: