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: iScience

Article Title: FOXE1 promotes the progression of pulp inflammation by activating PANoptosis in dental pulp cells

doi: 10.1016/j.isci.2026.115204

Figure Lengend Snippet: Identification of a PANoptotic DPC cluster in human clinical pulpitis samples (A) Gene sets associated with PANoptosis were constructed on the basis of previous studies. (B–D) The expression scores for pyroptosis-related (B, also see ), necroptosis-related (C, also see ), and apoptosis-related (D, also see ) genes were calculated using the AddModuleScore function in Seurat. NCs: neural cells, unknown: cells expressed mitochondrial genes as markers, RBCs: red blood cells, DPSCs: dental pulp stem cells, ECs: endothelial cells, GCs: glial cells, Ms: macrophages, TCs: T cells, DPCs: dental pulp cells, BCs: B cells, DCs: dendritic cells, PCs: plasma cells. (E) Multiple immunofluorescence staining of biomarkers of apoptosis, pyroptosis, and necroptosis in healthy and inflamed human dental pulp tissues. Vimentin-positive cells indicate DPCs. Scale bars, 10 μm. (F) Quantification of the staining results in E ( n = 4 samples in each group). Data are represented as mean ± SEM. ∗ p < 0.05 by Mann-Whitney U test (F).

Article Snippet: Annexin V-FITC/PI Apoptosis Kit , Multi Sciences , Cat#AP101.

Techniques: Construct, Expressing, Clinical Proteomics, Immunofluorescence, Staining, MANN-WHITNEY

Journal: iScience

Article Title: FOXE1 promotes the progression of pulp inflammation by activating PANoptosis in dental pulp cells

doi: 10.1016/j.isci.2026.115204

Figure Lengend Snippet: Mixed infection with S.m. and F.n. induced PANoptosis in DPCs (A–H) Dental pulp cells were infected with S.m. (MOI = 100), F.n. (MOI = 100), or a combination of S.m. (MOI = 100) and F.n. (MOI = 100) or treated with PBS (negative control, NC) for 8 h. (A) Dental pulp cells were treated with PBS, S.m. , F.n. , or a combination of S.m. and F.n. ( n = 5 experiments in each group), and cell death was quantified by measuring LDH release. (B) Dental pulp cells were treated with PBS, S.m. , F.n. , or a combination of S.m. and F.n . Treated DPCs were stained with PI, fixed, counterstained with DAPI, and visualized under a microscope. Scale bars, 100 μm. (C) Quantification of the proportions of PI-positive cells in B ( n = 6 experiments in each group). (D) Dental pulp cells were treated as indicated and analyzed by flow cytometry. (E) PANoptotic DPCs were quantified by determining the percentages of PI + /annexin V + cells ( n = 3 experiments in each group). (F) The protein levels of cleaved caspase-3 p17, cleaved GSDMD NT, and p -MLKL were assessed by Western blot. (G) Immunofluorescence staining of PANoptotic markers in infected DPCs, as visualized using a confocal microscope. Scale bars, 10 μm. (H) Quantification of PANoptotic cell proportions in DPCs treated with PBS, S.m. , F.n. , or a combination of S.m. and F.n. ( n = 8 samples in each group), corresponding to (G). Data are represented as mean ± SEM. ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by one-way ANOVA (A, C, and E) or by Kruskal-Wallis test (H).

Article Snippet: Annexin V-FITC/PI Apoptosis Kit , Multi Sciences , Cat#AP101.

Techniques: Infection, Negative Control, Staining, Microscopy, Flow Cytometry, Western Blot, Immunofluorescence

Journal: iScience

Article Title: FOXE1 promotes the progression of pulp inflammation by activating PANoptosis in dental pulp cells

doi: 10.1016/j.isci.2026.115204

Figure Lengend Snippet: Identification and validation of FOXE1 as a key transcription factor that regulates PANoptosis in DPCs (A) Differentially expressed gene analysis was performed between the PANoptotic DPC cluster (cluster 1) and other DPC clusters (clusters 0, 2, 4, 5, and 6) and between cluster 1 and all remaining clusters. The candidate regulators were selected from the overlapping DEGs between these comparisons. (B) The top five candidate regulators were identified after comparison. (C) siRNA targeting FOXE1 (20 μM) was transfected into DPCs with Lipofectamine 2000 for 24 h. The knockdown efficiency was validated by qRT-PCR ( n = 3 experiments in each group) and Western blot. (D–H) Dental pulp cells were pretreated with siRNAs (20 μM) targeting candidate regulators for 24 h, followed by infection with S.m. (MOI = 100) and F.n. (MOI = 100) for 8 h. (D) Cell death was quantified by the LDH release assays ( n = 5 experiments in each group). (E, also see in ) Cell death was assessed by PI staining ( n = 4 experiments in each group). (F) Treated cells were analyzed by flow cytometry. (G) PANoptotic DPCs were quantified by determining the percentages of PI + /annexin V + cells ( n = 3 experiments in each group). (H) Western blot analysis was used to assess the expression of cleaved caspase-3 p17, cleaved GSDMD NT and p -MLKL. (I) Immunofluorescence staining of PANoptotic markers in si FOXE1 -and siCON-transfected DPCs. Scale bars, 10 μm. (J) Quantification of PANoptotic DPCs in I ( n = 5 samples in each group). Data are represented as mean ± SEM. ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by Student’s t test (C, G, and J) or by one-way ANOVA (D and E).

Article Snippet: Annexin V-FITC/PI Apoptosis Kit , Multi Sciences , Cat#AP101.

Techniques: Biomarker Discovery, Comparison, Transfection, Knockdown, Quantitative RT-PCR, Western Blot, Infection, Staining, Flow Cytometry, Expressing, Immunofluorescence