Journal: Journal of Sport and Health Science

Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

doi: 10.1016/j.jshs.2025.101111

Figure Lengend Snippet: Relative HSP abundances in whole skeletal muscle homogenates from young adults and older adults pre and post HIT exercise. Representative Westen blots of (A) HSP72, HSP27, and αB-crystallin and (B) phosphorylated HSP27 Ser15 (pHSP27 Ser15) and pαB-crystallin Ser59 in whole muscle homogenates from the vastus lateralis of the same individuals. Calibration curves of mixed muscle homogenates are indicated and were used to determine the relative number of given proteins (see Methods). Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Relative abundances of (C) HSP72, (D) HSP27, (E) pHSP27 Ser15, (F) αB-crystallin, and (G) pαB-crystallin Ser59 from young (circle) and older adults Pre (square) and older adults Post (triangle) HIT exercise are shown relative to average Old (pre) on a given gel (data are presented as mean ± SD). Individuals indicated by the number of symbols ( n : 5–7), with the same color assigned to the same individual and consistent across all graphs. * p ≤ 0.05 indicates Brown-Forsye and Welch’s and post hoc analysis using Games-Horwell. HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

Article Snippet: Details of antibodies used are as follows: HSP72 (1 in 500 mouse monoclonal, SMC100A; StressMarq Biosciences, Victoria, Canada); HSP27 (1 in 1000 mouse monoclonal, G3.1 ab2790; Abcam, Cambridge, UK); pHSP27 Ser15 (1 in 2000 monoclonal rabbit, ab76313; Abcam), pHSP27 Ser82 (1 in 2000 polyclonal mouse, ADI-SPA-524; Enzo Biochem, Farmingdale, NY, USA), αB-crystallin (1 in 1000 mouse monoclonal, SPA-222; StressGen Biotechnologies), pαB-crystallin Ser59 (1 in 1000 rabbit polyclonal, SPA-227; StressGen Biotechnologies).

Techniques: Staining, Membrane

Journal: Journal of Sport and Health Science

Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

doi: 10.1016/j.jshs.2025.101111

Figure Lengend Snippet: HSP abundances in type I and II skeletal muscle fibers from young and older adults. (A, C, and F) The MHC isoform present was determined in individual muscle fiber segments from the vastus lateralis and, following pooling into type I and type II groups from a given biopsy, were analyzed by Westen blotting. Westen blots of (A) HSP72, (C) HSP27 and pHSP27 Ser15, (F) αB-crystallin and pαB-crystallin Ser59, with MHC isoforms in groups of fibers. Stain-free gels are indicative of total protein loading, and molecular weights are indicated by markers collected under white light capture without moving the membrane between that and chemiluminescence detection. Calibration curves of mixed muscle homogenates are indicated. Relative protein abundances of (B) HSP72, (D) HSP27, (E) pHSP27 Ser15, (G) αB-crystallin, and (H) pαB-crystallin Ser59 in fibers from young (circle) and older adults (square) type I fibers (no outline) and type II fibers (outline). All fibers are expressed relative to the average older adult’s type I fibers. The same color is assigned to the same individual and is consistent with (data are presented as mean ± SD). * p < 0.05 and ** p < 0.01, mixed effect model Univariant using either Tukey’s or Games-Horwell’s multiple comparison test (see Methods). HIT = high-intensity training; HSP = heat shock protein; MHC = myosin heavy chain; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

Article Snippet: Details of antibodies used are as follows: HSP72 (1 in 500 mouse monoclonal, SMC100A; StressMarq Biosciences, Victoria, Canada); HSP27 (1 in 1000 mouse monoclonal, G3.1 ab2790; Abcam, Cambridge, UK); pHSP27 Ser15 (1 in 2000 monoclonal rabbit, ab76313; Abcam), pHSP27 Ser82 (1 in 2000 polyclonal mouse, ADI-SPA-524; Enzo Biochem, Farmingdale, NY, USA), αB-crystallin (1 in 1000 mouse monoclonal, SPA-222; StressGen Biotechnologies), pαB-crystallin Ser59 (1 in 1000 rabbit polyclonal, SPA-227; StressGen Biotechnologies).

Techniques: Staining, Membrane, Comparison

Journal: Journal of Sport and Health Science

Article Title: Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals

doi: 10.1016/j.jshs.2025.101111

Figure Lengend Snippet: HSP abundances in type I and II skeletal fibers from older adults pre- and post HIT exercise. Relative protein abundances of (A and B) HSP72, (C and D) HSP27, (E and F) pHSP27 Ser15, (G and H) αB-crystallin, and (I and J) pαB-crystallin Ser59 in fibers from old pre and old post HIT exercise. All fibers are expressed relative to the average old pre type I fibers or relative pre type II depending on fiber type. The same color is assigned to the same Individual, consistent in both graphs and all figures. * p < 0.05 and ** p < 0.01 indicated significant difference in paired t -test (except pHSP27 Ser15 Wilcoxon match-pair rank test). Representative blots are shown in . HIT = high-intensity training; HSP = heat shock protein; pαB-crystallin Ser59 = phospho-αB-crystallin at Serine59; pHSP27 Ser15 = phospho-HSP27 at Serine15.

Article Snippet: Details of antibodies used are as follows: HSP72 (1 in 500 mouse monoclonal, SMC100A; StressMarq Biosciences, Victoria, Canada); HSP27 (1 in 1000 mouse monoclonal, G3.1 ab2790; Abcam, Cambridge, UK); pHSP27 Ser15 (1 in 2000 monoclonal rabbit, ab76313; Abcam), pHSP27 Ser82 (1 in 2000 polyclonal mouse, ADI-SPA-524; Enzo Biochem, Farmingdale, NY, USA), αB-crystallin (1 in 1000 mouse monoclonal, SPA-222; StressGen Biotechnologies), pαB-crystallin Ser59 (1 in 1000 rabbit polyclonal, SPA-227; StressGen Biotechnologies).

Techniques:

Journal: Bioactive Materials

Article Title: GSH-responsive nanovaccine triggers immunogenic cell death and potent memory T cell immunity for durable, recurrence-free tumor eradication

doi: 10.1016/j.bioactmat.2026.04.025

Figure Lengend Snippet: In vitro evaluation of CDT-induced immunogenic cell death (ICD), dendritic cell (DC) maturation, and T cell activation . ( A ) Schematic illustration of CDT-triggered ICD and associated damage-associated molecular pattern (DAMP) release. ( B, C ) Bioluminescence images and quantitative analysis of extracellular ATP release from 4T1 cells after different treatments. ( D, E ) Quantitative analysis and CLSM images of HMGB1 release. ( F ) CLSM images showing CRT surface exposure in 4T1 cells after different treatments. ( G ) Schematic illustration of the co-culture system used to evaluate DC maturation and subsequent T cell activation. Immature DCs (iDCs) were isolated from bone marrow, and CD8 + T cells were isolated from the lymph nodes and spleens of naïve BALB/c mice. ( H, I ) Representative flow cytometric plots and quantitative analysis of mature DCs, defined as CD11c + CD80 + D86 + cells, after 24 h incubation with differently treated 4T1 cells, showing enhanced DC maturation in the SHINE group. ( J, K ) Representative flow cytometric plots and quantification of CD69 + cells among CD8 + T cells. ( L, M ) Representative flow cytometric plots and quantification of CD25 + cells among CD8 + T cells. ( N, O ) Quantification of granzyme B and perforin expression in CD8 + T cells, indicating enhanced T cell activation and effector function following SHINE treatment. ( P ) Schematic illustration of the in vitro tumor-killing assay. CDT-pretreated 4T1-Luc cells (CDT only) and CDT-pretreated 4T1-Luc cells co-cultured with effector CD8 + T cells at an effector-to-target (E:T) ratio of 5:1 were used to evaluate the additional contribution of immunotherapy. After 24 h, D-luciferin (150 μg mL −1 ) was added to quantify live tumor cells by luciferase activity. ( Q, R ) Bioluminescence images and quantitative analysis of surviving 4T1-Luc cells after treatment with CDT alone or combined CDT + immunotherapy at different concentrations, showing that addition of immunotherapy further enhanced tumor cell killing. M, MnO 2 @PEG; MR, MnO 2 @R848; MP, MnO 2 @aPD-L1; SHINE, MnO 2 @R848@aPD-L1. Data are presented as mean ± SD ( n = 3 ; n = 4 for (R)). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Panels A, G, and P created with BioRender.com .

Article Snippet: Tumor cell viability was assessed by adding D-luciferin (Caliper Life Sciences, Hopkinton, Massachusetts, USA; 150 μg mL −1 ) and quantifying bioluminescence using an IVIS Lumina XR system (PerkinElmer, Inc., Waltham, Massachusetts, USA).

Techniques: In Vitro, Activation Assay, Co-Culture Assay, Isolation, Incubation, Expressing, Cell Culture, Luciferase, Activity Assay