igg1  (SouthernBiotech)

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: Non-coding RNA Research

Article Title: Exosomal miRNA-218–5p derived from low-passage dermal papilla cells modulates hair follicle growth and development

doi: 10.1016/j.ncrna.2026.01.004

Figure Lengend Snippet: Exosomes derived from low-passage DPCs regulated HFSC proliferation. (A) Indirect immunofluorescence showing ALPL and PCNA expression in low-passage (P1) and high-passage (P8) DPCs (scale bar = 50 μm). (B) RT-qPCR analysis of HF development-related gene expression in P1 and P8 DPCs (unpaired two-tailed t -test, n = 3). (C) TEM images of exosomes from P1 DPCs (DPC-Exos P1) and P8 DPCs (DPC-Exos P8). (D) NTA measurement of particle size of DPC-Exos P1 and DPC-Exos P8. (E) Western blot detection of exosome-specific proteins in DPC-Exos P1 and DPC-Exos P8. (F) CCK-8 assay evaluating HFSC proliferation after treatment with DPC-Exos from P1 and P8 (one-way ANOVA, n = 5). (G) Flow cytometry analysis of HFSC apoptosis after treatment with DPC-Exos from P1 and P8 (one-way ANOVA, n = 3). ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: The Annexin V-FITC Apoptosis Detection Kit (Vazyme, China, Cat No. A214) was employed, and apoptosis rates were analyzed via flow cytometry using a FACSAria SORP instrument (Becton Dickinson, USA).

Techniques: Derivative Assay, Immunofluorescence, Expressing, Quantitative RT-PCR, Gene Expression, Two Tailed Test, Western Blot, CCK-8 Assay, Flow Cytometry

Journal: International Journal of Pharmaceutics: X

Article Title: A pH-responsive dual-drug nanoplatform for stromal remodeling and enhanced chemotherapy via MMP3/TGF- β inhibition

doi: 10.1016/j.ijpx.2026.100489

Figure Lengend Snippet: In vitro therapeutic efficacy and synergy analysis of RPAE-QM in 4 T1 cells. (A) Cell viability of 4 T1 cells incubated with different formulations for 48 h determined by MTT assay. (B) The corresponding IC 50 values of the respective treatments. (C) Representative flow cytometry plots of Annexin V-FITC/PI staining for apoptosis analysis in 4 T1 cells. (D) Quantitative analysis of the total apoptotic rate. (E) Dose-response curves of free Que., free DM1, and their combination used for quantitative synergy determination. (F) The Combination Index (CI) plot as a function of Fraction affected (Fa) generated using the Chou-Talalay method; the reference line at CI = 1 indicates an additive effect, while CI < 1 indicates synergism. Data are presented as mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001.

Article Snippet: After 24 h, the cells were harvested and stained with Annexin V-FITC/PI Cell Apoptosis Detection Kit (Beyotime, China) for flow cytometer analysis.

Techniques: In Vitro, Drug discovery, Incubation, MTT Assay, Flow Cytometry, Staining, Generated

Journal: Poultry Science

Article Title: High genotoxicity of CRISPR/Cas9 versus limited efficacy of CRISPRi in chicken primordial germ cells

doi: 10.1016/j.psj.2026.106722

Figure Lengend Snippet: CRISPR/Cas9 induces DNA damage and apoptosis in PGCs. (A) Flow cytometry analysis 24 h after electroporation, quantifying the proportion of Annexin V + /PI + cells. The horizontal axis indicates PI and the vertical axis Annexin V. The upper-left quadrant (Annexin V + /PI + ) represents late apoptotic cells, and the lower-right quadrant (Annexin V + only) represents early apoptotic cells. Upper panels: results after electroporation with Cas9 + various gRNAs; lower panels: results with dCas9 + various gRNAs. (B) Bar graph of Annexin V + /PI + percentages across groups. Cas9 editing induced a highly significant increase in late apoptosis. (C) γ-H 2 AX foci (green) detected by immunofluorescence 24 h after electroporation. Foci appear as discrete nuclear puncta; nuclei are counterstained with DAPI (blue). Scale bar = 10 µm. (D) Quantification of γ-H 2 AX foci per cell. Cas9 targeting resulted in a significant increase in γ-H 2 AX foci per cell, whereas dCas9 with sgRNA did not. Statistical significance determined by one-way ANOVA (p-values are indicated in the figure).

Article Snippet: After another centrifugation, the supernatant was removed and cells were resuspended in 500 μL of 1× Annexin V Binding Buffer (from Annexin V-FITC/PI Apoptosis Kit, Elabscience E-CK-A211).

Techniques: CRISPR, Flow Cytometry, Electroporation, Immunofluorescence

Journal: Poultry Science

Article Title: High genotoxicity of CRISPR/Cas9 versus limited efficacy of CRISPRi in chicken primordial germ cells

doi: 10.1016/j.psj.2026.106722

Figure Lengend Snippet: PGCs are more sensitive to DNA double-strand breaks than somatic cells. (A) Cell viability curves after 24 h treatment with increasing concentrations of etoposide (ETP; x-axis shows log 10 [ETP]). PGC viability dropped sharply even at low ETP doses, whereas CEF cells were more tolerant (viability data are mean ± SD of triplicates). (B) Apoptosis detection in PGCs treated with low-dose ETP. The proportion of Annexin V + /PI + cells (late apoptosis) was significantly elevated even at 0.03 µM ETP. (C) Western blot analysis of γ-H 2 AX protein in PGCs treated with varying ETP concentrations (left panel; β-actin as loading control). The bar graph (right) shows the ratio of γ-H 2 AX to β-actin band intensity, with a marked increase at 3 µM ETP (p-values are indicated in the figure). (D) Quantification of γ-H 2 AX foci per cell in THP-1 cells, male PGCs, and female PGCs after exposure to X-ray doses of 0, 2, 4, 6, and 8 Gy. After 48 h recovery, γ-H 2 AX foci increased significantly with higher radiation in THP-1, male PGCs, and female PGCs. Female PGC data points are red squares; male PGCs are blue squares; THP-1 are black circles. (E) Cell-cycle distribution of male vs. female PGCs after DNA damage. PGCs were irradiated (2, 4, 6 Gy), cultured 48 h, and analyzed by flow cytometry for cell-cycle phase (propidium iodide staining). Stacked bars show the percentage of cells in G 0 G 1 , S, and G 2 /M phases in untreated vs. irradiated cells. After damage, female PGCs predominantly arrested in G 2 /M (increased G 2 fraction), whereas male PGCs accumulated in S phase. Statistical significance by one-way ANOVA (p-values are indicated in the figure).

Article Snippet: After another centrifugation, the supernatant was removed and cells were resuspended in 500 μL of 1× Annexin V Binding Buffer (from Annexin V-FITC/PI Apoptosis Kit, Elabscience E-CK-A211).

Techniques: Western Blot, Control, Irradiation, Cell Culture, Flow Cytometry, Staining

Journal: Journal of Translational Autoimmunity

Article Title: Anti-IgLON5 encephalitis is associated with anti-retinal immunological reactivity without retinal alteration

doi: 10.1016/j.jtauto.2026.100359

Figure Lengend Snippet: Indirect immunofluorescence on sections of monkey retina. The fluorescence observed is identified by a black arrow. (A) Patient 4 (IgG4); (B) Patient 1 (IgG4); (C) Patient 3 (IgG1); (D) Patient 5 (IgG4); (E) Patient 2 (IgG1); (F) Patient 1 (IgG4) after immunoadsorption of IgLON5 antibodies; (G) Control with macular edema (IgG1); (H) Control with anti-Hu encephalitis (IgG1); (I) Control with CAR syndrome (IgG4). The different layers of the retina are identified by their initials: pigment epithelium (pe), photoreceptor layer (pr), outer grain layer (og), outer plexiform layer (op), inner grain layer (ig), inner plexiform layer (ip), ganglion cell layer (gc), nerve fibre layer (nf).

Article Snippet: The reactivity of the patients’ sera and/or CSF against the retina was evaluated retrospectively with frozen samples (−80°, EXPLAINEUR biobank) through an indirect immunofluorescence technique using sections of monkey retina (Ref. FA1172-1005, Euroimmun), detected with an FITC-labelled secondary antibody anti-human IgAGM (Euroimmun conjugate) or directed against IgA (ref. F0204, DAKO), IgM (ref. F0203, DAKO), IgG1 (ref. 9052-02, Southern Biotech) and IgG4 (ref. 9200-02, Southern Biotech).

Techniques: Immunofluorescence, Fluorescence, Control

Journal: Journal of Translational Autoimmunity

Article Title: Anti-IgLON5 encephalitis is associated with anti-retinal immunological reactivity without retinal alteration

doi: 10.1016/j.jtauto.2026.100359

Figure Lengend Snippet: Indirect immunofluorescence on sections of monkey retina. The fluorescence observed is identified by a black arrow. (A) Patient 4 (IgG4); (B) Patient 1 (IgG4); (C) Patient 3 (IgG1); (D) Patient 5 (IgG4); (E) Patient 2 (IgG1); (F) Patient 1 (IgG4) after immunoadsorption of IgLON5 antibodies; (G) Control with macular edema (IgG1); (H) Control with anti-Hu encephalitis (IgG1); (I) Control with CAR syndrome (IgG4). The different layers of the retina are identified by their initials: pigment epithelium (pe), photoreceptor layer (pr), outer grain layer (og), outer plexiform layer (op), inner grain layer (ig), inner plexiform layer (ip), ganglion cell layer (gc), nerve fibre layer (nf).

Article Snippet: The reactivity of the patients’ sera and/or CSF against the retina was evaluated retrospectively with frozen samples (−80°, EXPLAINEUR biobank) through an indirect immunofluorescence technique using sections of monkey retina (Ref. FA1172-1005, Euroimmun), detected with an FITC-labelled secondary antibody anti-human IgAGM (Euroimmun conjugate) or directed against IgA (ref. F0204, DAKO), IgM (ref. F0203, DAKO), IgG1 (ref. 9052-02, Southern Biotech) and IgG4 (ref. 9200-02, Southern Biotech).

Techniques: Immunofluorescence, Fluorescence, Control