Journal: Microbiology Spectrum

Article Title: Wine Yeast Cells Acquire Resistance to Severe Ethanol Stress and Suppress Insoluble Protein Accumulation during Alcoholic Fermentation

doi: 10.1128/spectrum.00901-22

Figure Lengend Snippet: Wine and laboratory yeast cells responded similarly to ethanol under laboratory conditions. Cells were prepared and exposed to stress, as described in the legend of Fig. 1 . The expression levels of Hsp70 (white bars) and Hsp104 (black bars) were assayed using Western blotting. Each value is expressed as the mean ± SE of the fold changes in the protein levels relative to cells without stress treatment ( n = 3).

Article Snippet: Hsp104 and Hsp70 protein levels were monitored by Western blotting using an anti-Hsp104 (ADI-SPA-1040-D; Enzo Life Sciences, Inc., Farmingdale, NY, USA), anti-Hsp70 (SMC-162c; StressMarq Biosciences, Inc., Victoria, BC, Canada), anti-mouse IgG, horseradish peroxidase (HRP)-linked (7076S; Cell Signaling Technology, Beverly, MA, USA), and anti-rabbit IgG, HRP-linked (7074S; Cell Signaling Technology) antibodies.

Techniques: Expressing, Western Blot

Journal: Microbiology Spectrum

Article Title: Wine Yeast Cells Acquire Resistance to Severe Ethanol Stress and Suppress Insoluble Protein Accumulation during Alcoholic Fermentation

doi: 10.1128/spectrum.00901-22

Figure Lengend Snippet: Hsp70 and Hsp104 levels in wine yeast cells during fermentation were assayed using Western blotting. Samples were prepared as described in the legend of Fig. 3 . As controls, log-phase cells cultured in SD medium were treated without stress (E0), exposed to 10% (vol/vol) ethanol for 180 min (E10), or pretreated with 6% ethanol for 180 min and then exposed to 10% (vol/vol) ethanol for 180 min (E6/10) in SD medium. Each value is expressed as the mean ± SE of fold changes in protein levels relative to E0.

Article Snippet: Hsp104 and Hsp70 protein levels were monitored by Western blotting using an anti-Hsp104 (ADI-SPA-1040-D; Enzo Life Sciences, Inc., Farmingdale, NY, USA), anti-Hsp70 (SMC-162c; StressMarq Biosciences, Inc., Victoria, BC, Canada), anti-mouse IgG, horseradish peroxidase (HRP)-linked (7076S; Cell Signaling Technology, Beverly, MA, USA), and anti-rabbit IgG, HRP-linked (7074S; Cell Signaling Technology) antibodies.

Techniques: Western Blot, Cell Culture

Journal: Frontiers in Physiology

Article Title: Hypoxic Induced Decrease in Oxygen Consumption in Cuttlefish ( Sepia officinalis ) Is Associated with Minor Increases in Mantle Octopine but No Changes in Markers of Protein Turnover

doi: 10.3389/fphys.2017.00344

Figure Lengend Snippet: Metabolite and protein levels in mantle, gill, and heart of Sepia officinalis under either normoxic (open bars) or hypoxic (closed bars) (50% dissolved oxygen saturation, 1 h) conditions. (A) octopine; (B) mantle free glucose and glycogen; (C) HSP70; (D) polyubiquitinated proteins. Statistical significance for octopine, HSP70, and polyubiquitinated proteins, was assessed with a 1-way ANOVA and for differences between glucose or glycogen levels with a t -test. N = 6 for all conditions except for free glucose in hypoxic mantle where N = 4. Differences between means or grouped means represent statistical difference (Tukey's multiple comparison test; P < 0.001). No differences were found in mantle free glucose and glycogen nor polyubiquitinated proteins ( P > 0.05).

Article Snippet: Levels of HSP70 were quantified by loading known quantities of a recombinant HSP70 (Enzo Life Sciences, #ADI-SPP-758, standard levels of 15, 30, and 60 ng per lane) along with the samples and using an HSP70 antibody (#AS05 083A, Agrisera).

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Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: In long-lasting cellular stress phases of melanoma cells, stress granules are dissolved by HSP70

doi: 10.1007/s00018-025-05939-8

Figure Lengend Snippet: Melanoma cells exhibit high levels of HSP70 and p-G3BP1. (a) Western blot analysis of phosphorylated G3BP1 (P-G3BP1, Ser149) in normal human epidermal melanocytes (NHEM; passages P5 and P6) and melanoma cell lines. Additional tumor cell lines including Panc-1 (pancreatic carcinoma), Hep3B and HepG2 (hepatocellular carcinoma), and HT29 and SW480 (colorectal carcinoma) were included for comparison. ACTINB served as a loading control. (b) Densitometric quantification of P-G3BP1/G3BP1 protein ratio in NHEM, SK-Mel-28, WM1366, Mel Ho, Mel Juso, Sk-Mel-3 and WM35 cells. NHEM protein expression was set to 1. Normalized to ACTINB. (c) RNA sequencing analysis of HSP70 paralogues (HSPA1A, HSPA1B) in melanoma cell lines and NHEM. Read counts were normalized to library size. (d) Quantitative real-time PCR of HSPA1A and HSPA1B mRNA levels in melanoma cell lines and NHEM. (e) Western blot analysis of total HSP70 protein expression in NHEM (passages P5 and P8) and melanoma cell lines. ACTINB served as a loading control. (f) Western blot analysis of p-G3BP1 expression following treatment of Mel Juso and Mel Im cells with hypoxia-mimetic agents Desferrioxamine (DFX) and Dimethyloxalylglycine (DMOG). ACTINB was used as a loading control. (g) Western blot analysis of P-G3BP1 and G3BP1 expression in Mel Juso cells exposed to hypoxic conditions (1% O₂). ACTINB served as loading controls. (h) Western blot analysis of HSP70 expression in Mel Juso and Mel Im cells treated with DFX and DMOG. ACTINB served as a loading control. (i) Western blot analysis of HSP70 expression in Mel Juso cells under hypoxic conditions (1% O₂). ACTINB was used as a loading control

Article Snippet: In parallel, we observed that melanoma cells express high levels of the molecular chaperone HSP70 under basal conditions, a feature confirmed through analysis of The Human Protein Atlas and our experimental datasets.

Techniques: Western Blot, Comparison, Control, Expressing, RNA Sequencing, Real-time Polymerase Chain Reaction

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: In long-lasting cellular stress phases of melanoma cells, stress granules are dissolved by HSP70

doi: 10.1007/s00018-025-05939-8

Figure Lengend Snippet: HSP70 suppresses stress granule (SG) formation and protects melanoma cells from stress-induced apoptosis. (a) Quantitative real-time PCR analysis of HSP70 (HSPA1A/B) mRNA levels in Mel Juso cells 24 h after transfection with an HSP70 siRNA pool (siHSP70). (b) Western blot analysis of HSP70 protein levels in Mel Juso cells following siHSP70 transfection for 24 h. ACTINB served as a loading control. (c) Western blot analysis of HSP70, phosphorylated EIF2S1 (P-EIF2S1), and total EIF2S1 in Mel Juso cells transfected with siHSP70 for 24 h and subsequently treated with 600 µM sodium arsenite (SA) to induce stress. EIF2S1 served as the internal control. (d) Immunofluorescence staining of G3BP1 (red) in Mel Juso cells transfected with either siHSP70 or non-targeting control siRNA (sictrl), followed by SA treatment. Arrows indicate SG formation. Quantification of G3BP1-positive cells is shown; data represent the mean of three independent experiments. (e) Western blot analysis of total G3BP1 and phosphorylated G3BP1 (P-G3BP1) in Mel Juso cells after transfection with siHSP70 or sictrl and SA treatment. ACTINB was used as a loading control. Densitometric quantification from three independent experiments is shown. (f) Western blot analysis of PARP/cleaved PARP and CASPASE9/cleaved CASPASE9 in Mel Juso cells transfected with siHSP70 or sictrl, followed by SA treatment. Full-length PARP and CASPASE9 served as loading controls. Densitometric analysis from three independent experiments is provided. (g) Flow cytometric analysis of apoptotic Mel Juso cells after transfection with siHSP70 or sictrl and subsequent SA treatment. Data are presented as mean ± SEM (* p < 0.05, ** p < 0.01 and *** p < 0.001)

Article Snippet: In parallel, we observed that melanoma cells express high levels of the molecular chaperone HSP70 under basal conditions, a feature confirmed through analysis of The Human Protein Atlas and our experimental datasets.

Techniques: Real-time Polymerase Chain Reaction, Transfection, Western Blot, Control, Immunofluorescence, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: In long-lasting cellular stress phases of melanoma cells, stress granules are dissolved by HSP70

doi: 10.1007/s00018-025-05939-8

Figure Lengend Snippet: Caseinkinase 2a regulates Stress Granule Formation by modulating G3BP1 phosphorylation (a) Analysis of CSNK2A1 expression in skin cutaneous melanoma (SKCM; T) samples (red) compared to non-tumor skin (N) samples (grey). Data were obtained from the GEPIA database with a p-value cut-off of 0.01 and |Log₂FC| cut-off of 1 (retrieved August 21, 2025). (b) Representative western blot analysis of casein kinase 2α (CK2α) expression in normal human epidermal melanocytes (NHEMs) from different donors and melanoma cell lines (SK-MEL-28, WM1366, SK-MEL-3, Mel Juso, Mel Ho, and WM35). ACTINB was used as a loading control. (c) Schematic representation of the experimental timeline for TBCA treatment. (d) Immunofluorescence staining of G3BP1 (red) in Mel Juso cells treated with TBCA or DMSO for 45 min (stress phase) or after 24 h (recovery phase). Nuclei were counterstained with DAPI (blue). Quantification of stress granule (SG)-positive cells per field of view is shown as the mean of three independent experiments (lower panels). (e) Representative western blot analysis of HSP70, phosphorylated G3BP1 (P-G3BP1), and CK2α in cells treated with TBCA or DMSO, followed by sodium arsenite (SA) treatment for 45 min (stress phase) or 24 h (recovery phase). ACTINB served as a loading control

Article Snippet: In parallel, we observed that melanoma cells express high levels of the molecular chaperone HSP70 under basal conditions, a feature confirmed through analysis of The Human Protein Atlas and our experimental datasets.

Techniques: Phospho-proteomics, Expressing, Western Blot, Control, Immunofluorescence, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: In long-lasting cellular stress phases of melanoma cells, stress granules are dissolved by HSP70

doi: 10.1007/s00018-025-05939-8

Figure Lengend Snippet: Role of HSP70 in vemurafenib-resistant melanoma cells. (a) Representative western blot analysis of HSP70 and CK2α protein expression in SK-Mel-28 non-resistant (NR) and vemurafenib-resistant (R) cells. ACTINB was used as a loading control. (b) Western blot analysis of HSP70 and phosphorylated EIF2S1 (p-EIF2S1) expression in SK-Mel-28-R cells transfected with HSP70 siRNA (siHSP70) or non-targeting control siRNA (sictrl) for 24 h. Total EIF2S1 and ACTINB were used as loading controls. Densitometric quantification from three independent experiments is shown. (c) Immunofluorescence staining of G3BP1 (red) in SK-Mel-28-R cells following sodium arsenite (SA) treatment (600 µM) for 24 h. Nuclei were counterstained with DAPI (blue). Quantification of SG-positive cells per field of view is shown as the mean of three independent experiments. (d) Western blot analysis of phosphorylated G3BP1 (p-G3BP1) in SK-Mel-28-R cells transfected with siHSP70 or sictrl, followed by SA treatment for 24 h. Total G3BP1 and ACTINB served as loading controls. Quantification from three independent experiments is included. (e) Western blot analysis of full-length and cleaved PARP and CASPASE9 in SK-Mel-28-R cells after transfection with siHSP70 or sictrl, with or without additional SA treatment. ACTINB was used as a loading control. (f) Flow cytometric analysis of apoptosis in SK-Mel-28-R cells after siHSP70 or sictrl transfection (24 h), followed by SA treatment. Data reflect one representative experiment of three independent replicates. Data are presented as mean ± SEM (* p < 0.05 and **** p < 0.0001; ns = not significant)

Article Snippet: In parallel, we observed that melanoma cells express high levels of the molecular chaperone HSP70 under basal conditions, a feature confirmed through analysis of The Human Protein Atlas and our experimental datasets.

Techniques: Western Blot, Expressing, Control, Transfection, Immunofluorescence, Staining

Journal: Cell & Bioscience

Article Title: Heat shock protein 70 protects mouse against post-infection irritable bowel syndrome via up-regulating intestinal γδ T cell’s Th17 response

doi: 10.1186/s13578-018-0237-z

Figure Lengend Snippet: Protein and mRNA expression of HSP 70. a , b Expression of HSP 70 detected by Western blot; c , d Expression of HSP 70 detected by RT-PCR. *p < 0.05 compared with control group; △, p < 0.05 compared with both HSP70 group and IP-IBS group

Article Snippet: HSP70 protein level was measured by Western blot (Wuhan Boster Coporation, Wuhan, China).

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Cell & Bioscience

Article Title: Heat shock protein 70 protects mouse against post-infection irritable bowel syndrome via up-regulating intestinal γδ T cell’s Th17 response

doi: 10.1186/s13578-018-0237-z

Figure Lengend Snippet: γδ T cells’ Th17 response and IL-17 production by γδ T cells. a Purified γδ T cells were stained with Anti-TCRγ/δ-FITC antibodies and runned on FACS; b γδ T cells’ proliferation index was measured; c Annexin V-FITC/PI apoptosis Kit was used to determine the apoptosis rate; d The surface molecular expression of CD62L and CD69 was detected by FACS; e The production of IL17 by γδ T cells stimulated by HSP70 was measured by ELISA

Article Snippet: HSP70 protein level was measured by Western blot (Wuhan Boster Coporation, Wuhan, China).

Techniques: Purification, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Cryoablation inhibition of distant untreated tumors (abscopal effect) is immune mediated

doi: 10.18632/oncotarget.24105

Figure Lengend Snippet: (A) The serum Hsp70 level changed with time. (B) The differences in serum Hsp70 levels among the three groups. (C) The cytoplasmic Hsp70 level in untreated tumors on days 7 and 14 after treatment.

Article Snippet: Total Hsp70 levels in serum samples of mice were measured using the Mouse Hsp70 ELISA Kit (Lot CSB-E08311m, Cusabio.

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