criterion tgx stain  (Bio-Rad)

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: Self-healing and characterization of GMI gel . A) CD spectrum of GM, MI, and GMI at a concentration of 0.1 mg/mL. B) Representative TEM image of GMI. C) Rheology of GMI gel under frequency sweeps in the range of 0.1 - 100 rad/s. D) Rheology of GMI gel as a function of time. E) Strain sweeps of GMI gel as function of shear strain. F) The self-healing analysis of GMI gel with an oscillating force (100%) alternating with a small one (5%). G) Representative SEM images of GMI gel .

Article Snippet: The interior morphology of GMI gel was examined by SEM (Hitachi S-4800, Tokyo, Japan).

Techniques: Concentration Assay, Shear

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: The antimicrobial capacity of GMI gel . A, B) Effect of GMI gel prepared by different proportions of MI on the survival rate of MRSA and E. coli , n = 5. C-E) Effects of different treatments on ATP, K + and β-GAL in MRSA, n = 3. F) Photographs of MRSA and E. coli colonies after different treatments. G) SEM images of MRSA and E. coli bacteria after different treatments. The yellow arrow indicates the disruption of the bacterial cell membrane. H) Representative TEM images of GMI gel -treated MRSA and E. coli. I) Live/dead staining fluorescence images of MRSA and E. coli after GMI gel treatment. J) Live/dead stained 3D fluorescence image of MRSA biofilm after GMI gel treatment. K, L) Representative images and quantitative analysis of MRSA biofilm crystal violet staining after different treatments, n = 3. Data are shown as mean ± SDs. ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The interior morphology of GMI gel was examined by SEM (Hitachi S-4800, Tokyo, Japan).

Techniques: Bacteria, Disruption, Membrane, Staining, Fluorescence

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: GMI gel alleviates oxidative stress damage in macrophages and improves their mitochondrial function. A, B) Representative fluorescence images and quantitative analysis of mitochondrial superoxide scavenging by GMI gel in RAW264.7 cells., n = 3. C) The JC-1 staining method is used to detect the MMP in RAW264.7 cells. D) Quantitative analysis of MMPs, n = 3. E) ATP levels in RAW264.7 cells after different treatments, n = 3. F) Representative transmission electron microscopy images of mitochondria in RAW264.7 cells after different treatments. Data are shown as mean ± SDs. ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns, not significant (p > 0.05).

Article Snippet: The interior morphology of GMI gel was examined by SEM (Hitachi S-4800, Tokyo, Japan).

Techniques: Fluorescence, Staining, Transmission Assay, Electron Microscopy

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: GMI gel -mediated metabolic reprogramming and its regulatory role in macrophage polarization. A-D) Hexokinase activity, Phosphofructokinase activity, Isocitrate dehydrogenase activity, and Succinate dehydrogenase activity, n = 3. E-H) Heatmap of LC-MS data and quantitative analysis of the relative abundance of glycolysis and TCA cycle metabolites, n = 3. I) Schematic diagram of glycolysis and the TCA cycle. J) Representative CLSM images of RAW 264.7 cells treated with GMI gel for 72 h. K) Flow cytometry analysis of CD206 and CD86 expression. L, M) The secretion of IL-6 and TNF-α in the culture supernatant of RAW264.7 cells, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns, not significant (p > 0.05).

Article Snippet: The interior morphology of GMI gel was examined by SEM (Hitachi S-4800, Tokyo, Japan).

Techniques: Activity Assay, Liquid Chromatography with Mass Spectroscopy, Flow Cytometry, Expressing

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: The biocompatibility, cell migration, and tube formation treated by GMI gel . A) 3D images of L929 cell growth in GMI gel . B-D) Cell viability of L929, RAW264.7 cells and HUVECs cultured in GMI gel , n = 3. E) Hemolysis rate statistics of GMI gel , n = 3. F, G) Cell migration images and quantitative analysis of HUVECs after co-culture with GMI gel , n = 3. H, I) Tube formation images and quantitative analysis of HUVECs cultured in GMI gel , n = 3. Data are shown as mean ± SDs. ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001, ns, not significant (p > 0.05).

Article Snippet: The interior morphology of GMI gel was examined by SEM (Hitachi S-4800, Tokyo, Japan).

Techniques: Migration, Cell Culture, Co-Culture Assay

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: GMI gel promotes the healing of infected pressure ulcers in vivo. A) Schematic diagram of GMI gel treatment of infected pressure ulcers. B) Photographs of wounds in mice at different treatment times. C) Signs of wound closure. D) Wound size at different treatment times, n = 3. E) H&E staining images of mouse wound tissue after different treatments on day 12. F) Masson staining images of mouse wound tissue after different treatments on day 12. G) Representative laser Doppler perfusion images of wounds in mice in each treatment group on day 12. H) Representative images of immunohistochemical staining for TNF- α, IL-6 and IL-10 12 days after treatment. I-L) Quantitative statistics of wound site blood perfusion, TNF- α, IL-6 and IL-10, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The interior morphology of GMI gel was examined by SEM (Hitachi S-4800, Tokyo, Japan).

Techniques: Infection, In Vivo, Staining, Immunohistochemical staining

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: GMI gel promotes vascular regeneration and regulates mitochondrial metabolism. A) Representative images of DHE immunofluorescence staining on treatment day 12. B) Representative images of CD68 and CD86 immunofluorescence staining on treatment day 12. C)Representative images of CD68 and CD206 immunofluorescence staining on treatment day 12. D)Representative images of CD31 and α-SMA immunofluorescence staining on treatment day 12. E-I) Quantified results of relative fluorescence intensity for DHE, CD86, CD206, CD31, and α-SMA, n = 3. J) Representative TEM images of macrophage mitochondrial structure at pressure sore wounds on day 12 of treatment. K-N) Quantitative analysis of HK, PFK, IDH and SDH activities on day 12 of treatment, n = 3. Data are shown as mean ± SDs. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The interior morphology of GMI gel was examined by SEM (Hitachi S-4800, Tokyo, Japan).

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: Bioactive Materials

Article Title: Energetic metabolism-regulatory glycopeptide hydrogel accelerates pressure ulcer wound repair

doi: 10.1016/j.bioactmat.2026.02.016

Figure Lengend Snippet: RNA sequencing of cutaneous wound healing by GMI gel (n=3). A) Venn diagram showing the numbers of unique RNAs among the control group (C), GMI gel group (T), and Normal group (N). B) Volcano map of gene expression in GMI gel treatment group compared with the control group. Red and blue points represent upregulated and downregulated genes selected based p-adj <0.05 and |logFC|>2, respectively. C) Cluster heatmap for DEGs among the control group (C), GMI gel group (T), and Normal group (N). D) GO enrichment analysis of DEGs. E) KEGG pathway enrichment analysis of DEGs. FC, fold change; DEGs, differentially expressed genes; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: The interior morphology of GMI gel was examined by SEM (Hitachi S-4800, Tokyo, Japan).

Techniques: RNA Sequencing, Control, Gene Expression

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: Total protein was separated on 10% or 4%–15% Criterion TGX stain-free gels (Bio-Rad Laboratories) and run for 45 min at 200 V. Using a wet transfer protocol, protein was transferred to nitrocellulose membranes at 100 V for 30 min. Membranes were incubated in Miser TM solution (ThermoFisher Scientific) and blocked in 5% skim milk powder in tris-buffered saline-tween (TBST).

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: Total protein was separated on 10% or 4%–15% Criterion TGX stain-free gels (Bio-Rad Laboratories) and run for 45 min at 200 V. Using a wet transfer protocol, protein was transferred to nitrocellulose membranes at 100 V for 30 min. Membranes were incubated in Miser TM solution (ThermoFisher Scientific) and blocked in 5% skim milk powder in tris-buffered saline-tween (TBST).

Techniques: Staining, Membrane, Comparison