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: Anti-Cancer Drugs

Article Title: EP4 influences bortezomib resistance in multiple myeloma by modulating endoplasmic reticulum stress via the phosphatidylinositol 3-kinase/protein kinase B pathway

doi: 10.1097/CAD.0000000000001792

Figure Lengend Snippet: EP4 overexpression suppresses proliferation and migration while enhancing apoptosis in multiple myeloma cells. (a) Validation of EP4 overexpression in NCI-H929 and RPMI 8226 cells by qRT-PCR and Western blot. (*** P < 0.001, NC: control group; EP4-OE: EP4 overexpression group). (b–d) Effects of EP4 overexpression on cell viability, proliferation, and apoptosis. (b) CCK-8 assay, (c) EdU staining, and (d) Annexin V-PI staining of apoptotic events. (e) GSEA analysis using the high and low- EP4 expression groups in GSE24080 dataset, showing correlation with endoplasmic reticulum membrane protein with high- EP4 expressing group. (f) Western blot analysis of GRP78 expression in control and EP4-overexpressing cells. (g, h) Transwell assays for cell migration and invasion in control and EP4-overexpressing cells. (** P < 0.01, *** P < 0.001). CCK-8, Cell Counting Kit-8; GSEA, gene set enrichment analysis; qRT-PCR, quantitative reverse transcription-PCR.

Article Snippet: The Annexin V-FITC/PI apoptosis kit was acquired from Hangzhou Lianke Biotechnology Co. (Hangzhou, Zhejiang, China).

Techniques: Over Expression, Migration, Biomarker Discovery, Quantitative RT-PCR, Western Blot, Control, CCK-8 Assay, Staining, Expressing, Membrane, Cell Counting, Reverse Transcription

Journal: Anti-Cancer Drugs

Article Title: EP4 influences bortezomib resistance in multiple myeloma by modulating endoplasmic reticulum stress via the phosphatidylinositol 3-kinase/protein kinase B pathway

doi: 10.1097/CAD.0000000000001792

Figure Lengend Snippet: EP4 overexpression enhances bortezomib sensitivity in resistant multiple myeloma cells. (a–c) Effects of EP4 overexpression combined with bortezomib treatment on (a) cell viability, (b) proliferation, and (c) apoptosis in OPM-2/BTZ and KMS-11/BTZ cells. (** P < 0.01, *** P < 0.001). (d, e) Western blot analysis of (d) GRP78 expression and (e) PI3K/AKT pathway activation in EP4-overexpressing resistant cells. All BTZ groups were treated with 10 nM bortezomib for 48 h. BTZ, bortezomib; PI3K/AKT, phosphatidylinositol 3-kinase/protein kinase B.

Article Snippet: The Annexin V-FITC/PI apoptosis kit was acquired from Hangzhou Lianke Biotechnology Co. (Hangzhou, Zhejiang, China).

Techniques: Over Expression, Western Blot, Expressing, Activation Assay

Journal: Anti-Cancer Drugs

Article Title: EP4 influences bortezomib resistance in multiple myeloma by modulating endoplasmic reticulum stress via the phosphatidylinositol 3-kinase/protein kinase B pathway

doi: 10.1097/CAD.0000000000001792

Figure Lengend Snippet: EP4 modulates bortezomib resistance through AKT-regulated endoplasmic reticulum stress. (a–c) Effects of EP4 overexpression, AKT agonist SC79, and ER stress inducer Thapsigargin on (a) cell viability, (b) apoptosis, and (c) GRP78 expression in OPM-2/BTZ and KMS-11/BTZ cells. All groups of cells were treated with 10 nM bortezomib for 48 h. (*** P < 0.001 vs. EP4-NC group, ### P < 0.001 vs. EP4-OE group, ^^^ P < 0.001 vs. EP4-OE+SC79 group). AKT, protein kinase B; ER, endoplasmic reticulum.

Article Snippet: The Annexin V-FITC/PI apoptosis kit was acquired from Hangzhou Lianke Biotechnology Co. (Hangzhou, Zhejiang, China).

Techniques: Over Expression, Expressing

Journal: Anti-Cancer Drugs

Article Title: EP4 influences bortezomib resistance in multiple myeloma by modulating endoplasmic reticulum stress via the phosphatidylinositol 3-kinase/protein kinase B pathway

doi: 10.1097/CAD.0000000000001792

Figure Lengend Snippet: Schematic illustration of EP4-mediated regulation of bortezomib resistance in multiple myeloma. EP4 expression decreases in bortezomib-resistant cells. EP4 overexpression restores sensitivity by inhibiting PI3K/AKT signaling, increasing ER stress, and promoting apoptosis. This mechanism suggests EP4 as a potential therapeutic target for overcoming bortezomib resistance. PI3K/AKT, phosphatidylinositol 3-kinase/protein kinase B.

Article Snippet: The Annexin V-FITC/PI apoptosis kit was acquired from Hangzhou Lianke Biotechnology Co. (Hangzhou, Zhejiang, China).

Techniques: Expressing, Over Expression