Journal: International Journal of Clinical and Experimental Pathology

Article Title: High expression of heat shock protein 10 (Hsp10) is associated with poor prognosis in oral squamous cell carcinoma

doi:

Figure Lengend Snippet: Expression of Hsp10 in OSCC tissue was detected by IHC. The expression of Hsp10 was detected by IHC using specific antibodies as described in the section of materials and methods. Strong positive staining of Hsp10 was identified in the cytoplasm of OSCC (A, 20×, IHC, DAB staining). Moderate positive staining of Hsp10 was showed in the cytoplasm of OSCC cells (B, 20×, IHC, DAB staining). Weak positive staining of Hsp10 was localized in the cytoplasm of OSCC cells (C, 20×, IHC, DAB staining). Negative staining of Hsp10 was in the OSCC cells (D, 20×, IHC, DAB staining).

Article Snippet: After incubation with a 1:1000 dilution of primary antibody to Hsp10 protein (Mouse monoclonal antibody, Catalog: sc-376313, Santa Cruz Biotechnology, Inc.) at 4°C overnight, slides washed with physiological phosphate buffered saline (PBS) three times for 10 minutes each and second antibody conjugated with a labeled polymer-HRP was added in accordance with the manufacturer’s instructions and incubated at 37°C for 30 minutes.

Techniques: Expressing, Staining, Negative Staining

Journal: International Journal of Clinical and Experimental Pathology

Article Title: High expression of heat shock protein 10 (Hsp10) is associated with poor prognosis in oral squamous cell carcinoma

doi:

Figure Lengend Snippet: Expression of Hsp10 in the non-cancerous squamous epithelium was detected by IHC. The expression of Hsp10 was detected by IHC using specific antibodies as described in the section of materials and methods. Moderate positive staining of Hsp10 was showed in the cytoplasm of the non-cancerous squamous epithelium (A, 20×, IHC, DAB staining). Weak positive staining of Hsp10 was displayed in the cytoplasm of the non-cancerous squamous epithelium (B, 20×, IHC, DAB staining).

Article Snippet: After incubation with a 1:1000 dilution of primary antibody to Hsp10 protein (Mouse monoclonal antibody, Catalog: sc-376313, Santa Cruz Biotechnology, Inc.) at 4°C overnight, slides washed with physiological phosphate buffered saline (PBS) three times for 10 minutes each and second antibody conjugated with a labeled polymer-HRP was added in accordance with the manufacturer’s instructions and incubated at 37°C for 30 minutes.

Techniques: Expressing, Staining

Journal: International Journal of Clinical and Experimental Pathology

Article Title: High expression of heat shock protein 10 (Hsp10) is associated with poor prognosis in oral squamous cell carcinoma

doi:

Figure Lengend Snippet: Association between expression of Hsp10 and OSCC clinical pathological feature (n = 110)

Article Snippet: After incubation with a 1:1000 dilution of primary antibody to Hsp10 protein (Mouse monoclonal antibody, Catalog: sc-376313, Santa Cruz Biotechnology, Inc.) at 4°C overnight, slides washed with physiological phosphate buffered saline (PBS) three times for 10 minutes each and second antibody conjugated with a labeled polymer-HRP was added in accordance with the manufacturer’s instructions and incubated at 37°C for 30 minutes.

Techniques: Expressing

Journal: International Journal of Clinical and Experimental Pathology

Article Title: High expression of heat shock protein 10 (Hsp10) is associated with poor prognosis in oral squamous cell carcinoma

doi:

Figure Lengend Snippet: Kaplan-Meier curves for overall survival of OSCC were assessed using the log-rank test. A: High expression of Hsp10 had been significantly related to short survival times of OSCC patients (P<0.001, two-sided). B: OSCC patients with well differentiation were significantly related to higher overall survival rates compared to patients with moderate and poor differentiation (P = 0.043, two-sided). C: OSCC patients with clinical stage III and IV were significantly related to lower overall survival rates compared to those patients with clinical stage I and II (P = 0.004, two-sided). D: OSCC patients with lymph node metastasis were significantly related to poor overall survival rates compared to those patients without lymph node metastasis (P = 0.033, two-sided).

Article Snippet: After incubation with a 1:1000 dilution of primary antibody to Hsp10 protein (Mouse monoclonal antibody, Catalog: sc-376313, Santa Cruz Biotechnology, Inc.) at 4°C overnight, slides washed with physiological phosphate buffered saline (PBS) three times for 10 minutes each and second antibody conjugated with a labeled polymer-HRP was added in accordance with the manufacturer’s instructions and incubated at 37°C for 30 minutes.

Techniques: Expressing

Journal: International Journal of Clinical and Experimental Pathology

Article Title: High expression of heat shock protein 10 (Hsp10) is associated with poor prognosis in oral squamous cell carcinoma

doi:

Figure Lengend Snippet: Summary of multivariate analysis of Cox proportional hazard regression for overall survival in 110 cases of OSCC patients

Article Snippet: After incubation with a 1:1000 dilution of primary antibody to Hsp10 protein (Mouse monoclonal antibody, Catalog: sc-376313, Santa Cruz Biotechnology, Inc.) at 4°C overnight, slides washed with physiological phosphate buffered saline (PBS) three times for 10 minutes each and second antibody conjugated with a labeled polymer-HRP was added in accordance with the manufacturer’s instructions and incubated at 37°C for 30 minutes.

Techniques: Expressing

Journal: Cell Stress & Chaperones

Article Title: Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer

doi: 10.1007/s12192-016-0721-5

Figure Lengend Snippet: Percentages of cells immunopositive for Hsps in large bowel mucosa

Article Snippet: The primary antibodies used were anti-human Hsp10 (rabbit polyclonal antibody, clone FL-102, Santa Cruz Biotechnology, Inc., Heidelberg, Germany, Europe, cat. no. Sc-28,887, dilution 1:200), anti-human Hsp60 (mouse monoclonal antibody, clone LK1, Sigma, St. Louis, MO, USA, Cat. No. H4149, dilution 1:400), anti-human Hsp70/HSC70 (mouse monoclonal antibody, clone W27, Santa Cruz Biotechnology, Inc., Europe, Cat. No. sc-24, dilution 1:200), and anti-human Hsp90 (mouse monoclonal antibody, clone F-8, Santa Cruz Biotechnology, Inc., Europe, Cat. No. sc-13,119, dilution 1:200).

Techniques:

Journal: Cell Stress & Chaperones

Article Title: Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer

doi: 10.1007/s12192-016-0721-5

Figure Lengend Snippet: Histograms show percentage of cells immunopositive for Hsp10, Hsp60, Hsp70, and Hsp90 in epithelial cells (EC) (a) and in cells of lamina propria (LPC) (b) in human large bowel normal mucosa (NM), tubular adenoma (TA), and adenocarcinoma (AC). Data are presented as the means ± SD. *p < 0.05

Article Snippet: The primary antibodies used were anti-human Hsp10 (rabbit polyclonal antibody, clone FL-102, Santa Cruz Biotechnology, Inc., Heidelberg, Germany, Europe, cat. no. Sc-28,887, dilution 1:200), anti-human Hsp60 (mouse monoclonal antibody, clone LK1, Sigma, St. Louis, MO, USA, Cat. No. H4149, dilution 1:400), anti-human Hsp70/HSC70 (mouse monoclonal antibody, clone W27, Santa Cruz Biotechnology, Inc., Europe, Cat. No. sc-24, dilution 1:200), and anti-human Hsp90 (mouse monoclonal antibody, clone F-8, Santa Cruz Biotechnology, Inc., Europe, Cat. No. sc-13,119, dilution 1:200).

Techniques:

Journal: Cell Stress & Chaperones

Article Title: Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer

doi: 10.1007/s12192-016-0721-5

Figure Lengend Snippet: Representative images of immunohistochemical results for Hsp10, Hsp60, Hsp70, and Hsp90 in human large bowel biopsies of normal mucosa, tubular adenoma with moderate grade of dysplasia, and adenocarcinoma with moderate grade of differentiation. Magnification 400×. Scale bar 100 μm. Arrows show the positivity for the Hsps in the epithelial cells of normal mucosa. Note the slight positivity for Hsp60 in normal epithelial cells

Article Snippet: The primary antibodies used were anti-human Hsp10 (rabbit polyclonal antibody, clone FL-102, Santa Cruz Biotechnology, Inc., Heidelberg, Germany, Europe, cat. no. Sc-28,887, dilution 1:200), anti-human Hsp60 (mouse monoclonal antibody, clone LK1, Sigma, St. Louis, MO, USA, Cat. No. H4149, dilution 1:400), anti-human Hsp70/HSC70 (mouse monoclonal antibody, clone W27, Santa Cruz Biotechnology, Inc., Europe, Cat. No. sc-24, dilution 1:200), and anti-human Hsp90 (mouse monoclonal antibody, clone F-8, Santa Cruz Biotechnology, Inc., Europe, Cat. No. sc-13,119, dilution 1:200).

Techniques: Immunohistochemical staining

Journal: Cell Stress & Chaperones

Article Title: Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer

doi: 10.1007/s12192-016-0721-5

Figure Lengend Snippet: Tubular adenoma. Representative high-power images of immunostaining for Hsp10 and Hsp60 in lamina propria cells with the typical characteristics of inflammatory cells (arrows). (Magnification 630× and 1000×)

Article Snippet: The primary antibodies used were anti-human Hsp10 (rabbit polyclonal antibody, clone FL-102, Santa Cruz Biotechnology, Inc., Heidelberg, Germany, Europe, cat. no. Sc-28,887, dilution 1:200), anti-human Hsp60 (mouse monoclonal antibody, clone LK1, Sigma, St. Louis, MO, USA, Cat. No. H4149, dilution 1:400), anti-human Hsp70/HSC70 (mouse monoclonal antibody, clone W27, Santa Cruz Biotechnology, Inc., Europe, Cat. No. sc-24, dilution 1:200), and anti-human Hsp90 (mouse monoclonal antibody, clone F-8, Santa Cruz Biotechnology, Inc., Europe, Cat. No. sc-13,119, dilution 1:200).

Techniques: Immunostaining

Journal: Cell Stress & Chaperones

Article Title: An inventory of interactors of the human HSP60/HSP10 chaperonin in the mitochondrial matrix space

doi: 10.1007/s12192-020-01080-6

Figure Lengend Snippet: Strategy for identification of interactors of the HSP60/HSP10 chaperonin. HEK293 cells were grown at 37 °C in light, medium, and heavy SILAC medium, respectively. One of the cultures in each experiment, [+HS], was treated at 43 °C for 3 h, followed by 12 h of recovery at 37 °C. At the end of incubation, cross-linker was added and cells were subsequently lysed and subjected to immunoprecipitation with antibodies directed against HSP60 (α-HSP60) or GFP (α-GFP) as indicated. The three immunoprecipitates were pooled and subjected to nano-LC-MS/MS analysis. The experiment was performed in three independent replicates with swapping of the media as indicated. Proteins quantified in at least 2 of the 3 experiments with a SILAC-ratio ≥ 3 between the HSP60 co-IPs versus the GFP co-IP in the respective experiments were classified as HSP60/HSP10 chaperonin interactors. Proteins quantitated in the HSP60 co-IP and having zero intensity in the GFP co-IP were also judged as fulfilling the SILAC ratio criterion. The interactors were cross-checked for presence in the MitoCarta 2.0 inventory of mitochondrial proteins, the inventory of mitochondrial matrix proteins, and the inventory of intermembrane space proteins

Article Snippet: Respective regions were cut from the membranes and incubated over night with primary antibodies directed against HSP60 (Thermo Scientific, cat. no. MA3–013) or HSP10 (Abnova, cat. no. {"type":"entrez-protein","attrs":{"text":"PAB15431","term_id":"1236628211","term_text":"PAB15431"}} PAB15431 ).

Techniques: Incubation, Immunoprecipitation, Liquid Chromatography with Mass Spectroscopy, Co-Immunoprecipitation Assay

Journal: Cell Stress & Chaperones

Article Title: An inventory of interactors of the human HSP60/HSP10 chaperonin in the mitochondrial matrix space

doi: 10.1007/s12192-020-01080-6

Figure Lengend Snippet: Overview of the interactors found. a Venn diagram showing the interactors of the HSP60/HSP10 chaperonin identified in the [−HS] co-IPs and assignment to the MitoCarta 2.0 (Calvo et al. 2016), mitochondrial matrix space (Rhee et al. 2013), and mitochondrial intermembrane space (IMS) inventories (Hung et al. 2014). (*) denotes that the protein numbers do not add up to the total number of proteins in the inventories because HSP60 (part of MitoCarta 2.0, and the matrix inventory) and HSP10 (part of all 3 inventories) are not included. b Table giving an overview over protein numbers in the mitochondrial protein inventories and the HSP60 interactors. The numbers of all interacting proteins found, proteins in the [−HS] and [+HS] co-IPs, respectively, and proteins common in both co-IPs are given

Article Snippet: Respective regions were cut from the membranes and incubated over night with primary antibodies directed against HSP60 (Thermo Scientific, cat. no. MA3–013) or HSP10 (Abnova, cat. no. {"type":"entrez-protein","attrs":{"text":"PAB15431","term_id":"1236628211","term_text":"PAB15431"}} PAB15431 ).

Techniques:

Journal: Cell Stress & Chaperones

Article Title: An inventory of interactors of the human HSP60/HSP10 chaperonin in the mitochondrial matrix space

doi: 10.1007/s12192-020-01080-6

Figure Lengend Snippet: Comparison of abundance of HSP60/HSP10 chaperonin interactors with cellular abundance. The log (10) iBAQ values of matrix assigned HSP60/HSP10 chaperonin interactors in the [−HS] co-IPs are plotted against their log (10) iBAQ values in HEK293 cell lysates (Geiger et al. 2012). The values for matrix proteins only identified in the [−HS] co-IP and not in the [+HS] co-IPs are depicted with gray squares and proteins identified in both the [+HS] and the [−HS] co-IPs are depicted with filled black circles. Squares and circles representing ribosomal subunits are marked with white crosses. Linear regression of all data points is shown by the stippled black line. Pearson’s correlation analysis showed a positive relationship with a correlation coefficient of 0.558. The log (10) iBAQ value corresponding to 1% occupancy of the HSP60/HSP10 chaperonin is indicated with an arrow on the horizontal axis. Points for the following subgroups of proteins and proteins referred to in the text are labeled as follows: the 19 proteins accounting for 63% of occupancy (black label on gray background); proteins with relatively high chaperonin occupancy in relation to their cellular abundance (boxed labels); additional proteins mentioned in the text (white label on black background)

Article Snippet: Respective regions were cut from the membranes and incubated over night with primary antibodies directed against HSP60 (Thermo Scientific, cat. no. MA3–013) or HSP10 (Abnova, cat. no. {"type":"entrez-protein","attrs":{"text":"PAB15431","term_id":"1236628211","term_text":"PAB15431"}} PAB15431 ).

Techniques: Comparison, Co-Immunoprecipitation Assay, Labeling

Journal: Cell Death & Disease

Article Title: Oxidative phosphorylation-dependent regulation of cancer cell apoptosis in response to anticancer agents

doi: 10.1038/cddis.2015.305

Figure Lengend Snippet: Etoposide induces mitochondrial unfolded protein response (UPR mt ). HCT116 cells were treated with etoposide (Etop; 10 μ M) for various time periods. ( a and b ) Whole cell lysates were prepared and subjected to western blotting for mentioned mitochondrial UPR proteins. Actin serves as loading control. ( c ) HCT116 cells treated with etoposide (Etop; 10 μ M) for 24 h. Equal amounts of total RNA were subjected to RNA-Seq library preparation followed by sequence reads (80 millions). Expression levels of CHOP, CEBP, ClpP, DnaJ, Hsp10, Hsp60, Hsp70, Hsp90, and HSF-2 are presented. ( d ) HCT116 cells treated with etoposide (Etop; 10 μ M) for 24 h. Total RNAs were isolated from treated and untreated cells and equal amounts of RNA was used for real-time PCR analysis for mentioned UPR genes including the expression levels of CHOP, CEBP, ClpP, DnaJ, Hsp10, Hsp60, Hsp70, Hsp90, and HSF-2. Data are mean±S.D., n =3

Article Snippet: The primary antibody against heat-shock protein (Hsp) 60 (Merck Millipore, Billerica, MA, USA), Hsp10 and Hsp70 (Enzo Life Sciences, Farmingdale, NY, USA), Hsp90 (R&D System, Minneapolis, MN, USA), ClpP (Abcam, Cambridge, MA, USA), and actin (mAb; ICN) were obtained from the indicated suppliers.

Techniques: Western Blot, RNA Sequencing Assay, Sequencing, Expressing, Isolation, Real-time Polymerase Chain Reaction