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

Journal: Bioactive Materials

Article Title: Glucose/ROS-responsive and redox-gated adaptive hydrogel dressing for accelerating diabetic wound repair via synergistic cGAS/STING pathway inhibition and oxidative stress alleviation

doi: 10.1016/j.bioactmat.2026.03.025

Figure Lengend Snippet: The cellular uptake and anti-inflammatory effect of HPSL in vitro . (A) Flow cytometry analysis and (B) semi-quantitative analysis of cellular uptake of PSL and blank NPs by M1 macrophages. n = 3. (C) Representative Giemsa staining images of LPS and high glucose-stimulated RAW 264.7 cells with different formulations, scale bar = 50 μm. (D) Immunofluorescence staining and semi-quantitative analysis of CD68 (red) and iNOS (green) in RAW 264.7 cells from different treatment groups, scale bar = 50 μm. n = 6. (E) Immunofluorescence staining and semi-quantitative analysis of CD68 (green) and Arg-1 (red) in RAW 264.7 cells from different treatment groups, scale bar = 50 μm. n = 6. Western blotting analysis and corresponding semi-quantitative analysis of (F) STING/ p -STING, (G) TBK1/ p -TBK1, (H) IRF3/ p -IRF3, (I) NF-κB, (J) TNF-α, and (K) IL-6, Lane 1: Normal group, Lane 2: Model group, Lane 3: PSL group, Lane 4: Free H151 group, Lane 5: HPSL group. n = 3. All data are shown as mean ± SEM.

Article Snippet: CD68-specific antibodies were purchased from Proteintech Group, Inc. (Wuhan, China).

Techniques: In Vitro, Flow Cytometry, Staining, Immunofluorescence, Western Blot

Journal: Bioactive Materials

Article Title: Inhalable PD-L1-engineered hybrid cellular vesicles suppress excessive neutrophil activation and restore mitochondrial homeostasis to alleviate ischemia–reperfusion lung injury and pneumonia

doi: 10.1016/j.bioactmat.2026.03.024

Figure Lengend Snippet: Res-PD-L1@nmEVs Attenuate Inflammation and Oxidative Damage in Lung Epithelial Cells In Vitro . (A-B) Flow cytometric analysis and quantification (B) of DiO-labeled Res-PD-L1@nmEVs uptake by BEAS-2B cells under H/R conditions after pretreatment with different endocytic inhibitors (chlorpromazine, chloroquine, and filipin) or incubation at 4 °C. (C) mRNA expression levels of IL-6, TNF-α, and IL-1β in BEAS-2B cells with or without H/R injury following pretreatment with Res, nEVs, PD-L1@mEVs, PD-L1@nmEVs, or Res-PD-L1@nmEVs. (D-E) Representative fluorescence images (D) and quantitative analysis (E) of cell proliferation assessed by BrdU incorporation (red; nuclei stained with DAPI, blue). Scale bar: 50 μm. (F-G) Apoptosis rates detected by flow cytometry (F) and flow cytometric analysis of Annexin V-positive BEAS-2B cells under the indicated conditions (G). (H–K) Fluorescence microscopy images and quantitative analysis of intracellular nitric oxide (NO, green) (H-I) and reactive oxygen species (ROS, red) (J-K). Scale bar: 100 μm. (L) Flow cytometry analysis of intracellular ROS levels. (M − O) Levels of malondialdehyde (MDA) (M), superoxide dismutase 2 (SOD2) activity (N), and glutathione (GSH) content (O) in cells. (P-Q) Cell migration ability evaluated by wound healing assay under different treatments. ∗ vs. Control; # vs. H/R; & vs. H/R + PD-L1@nmEVs, p < 0.05.

Article Snippet: Apoptosis was detected using double staining with Annexin V-APC and 7-AAD (35-640-KIT, Tonbo, USA) to distinguish early and late apoptotic cell populations.

Techniques: In Vitro, Labeling, Incubation, Expressing, Fluorescence, BrdU Incorporation Assay, Staining, Flow Cytometry, Microscopy, Activity Assay, Migration, Wound Healing Assay, Control

Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: MMP12 silencing inhibited M2 macrophage polarization. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. (A) Flow cytometry was used to quantify the number of CD68-positive cells. Subsequently, KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (B) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (C) Flow cytometry was used to quantify the number of CD206-positive macrophages. (D) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: Additionally, to determine the proportion of CD206-positive macrophages, single-cell suspensions of these cells were incubated with a FITC-conjugated anti-CD206 antibody (E-AB-F1161E, Elabscience).

Techniques: Flow Cytometry, Cell Culture, Derivative Assay, Quantitative RT-PCR, Migration, Transwell Migration Assay

Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: WTAP silencing inhibited M2 macrophage polarization by regulating MMP12. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. KYSE150 cells were transfected with si-WTAP, MMP12 overexpression plasmid, or the matched control (si-NC and oe-NC). Subsequently, these KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (A) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (B) Flow cytometry was used to quantify the number of CD206-positive macrophages. (C) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: Additionally, to determine the proportion of CD206-positive macrophages, single-cell suspensions of these cells were incubated with a FITC-conjugated anti-CD206 antibody (E-AB-F1161E, Elabscience).

Techniques: Transfection, Over Expression, Plasmid Preparation, Control, Cell Culture, Derivative Assay, Quantitative RT-PCR, Flow Cytometry, Migration, Transwell Migration Assay

Journal: International Journal of Oncology

Article Title: Diverse roles of SERPINE1 in regulating cellular proliferation and invasion

doi: 10.3892/ijo.2026.5871

Figure Lengend Snippet: SERPINE1-mediated modulation of the ERK/p38 ratio and anoikis. (A) Upper panel: western blotting showing ERK, p-ERK, p38, and p-p38 levels. Quantitative data showed the p-ERK/ERK and p-p38/p38 ratios in SERPINE1 knockdown cells compared with control cells. Lower panel: western blotting analysis of p-AKT, AKT, p-JNK and JNK (normalized to total protein) levels in the shSE1 and shc groups. The numbers beneath the bands represent the ratios of the expression levels of the indicated proteins. Data represent mean ± SDs of three independent experiments. (B) PLAUR and HSP90AA1 mRNA and protein levels in shSE1 and shc cells analyzed using RNA-seq (left panel) and western blotting (right panel). The numbers beneath the bands represent the ratios of the expression levels of the indicated proteins and represent mean ± SDs of three independent experiments. RNA-seq was performed using one biological replicate per group. (C-D) Upper panel: (C) 231-shSE1 cells were infected with lentivirus containing PLAUR cDNA (ex-PLAUR) or the corresponding empty vector control (vec). (D) H4-shSE1 cells were transfected with siRNA targeting HSP90AA1 (si-HSP90AA1) or siNC. The expression levels of uPAR and HSP90α, as well as the activity of ERK and p38, were detected by western blotting, with GAPDH serving as the loading control. Data represent mean ± SDs of three independent experiments. Lower panel: Cell proliferation was assessed by CCK-8 assay in 231-shSE1 cells infected with ex-PLAUR or vec, and in H4-shSE1 cells transfected with si-HSP90AA1 or siNC. Statistical significance was determined using two-way ANOVA followed by Šídák's multiple comparisons test. The data are presented as the means ± SDs. n=6. (E) Morphology of shSE1 and shc cells cultured under suspension condition. (scale bar, 100 μ m). (F) Apoptotic cells cultured under suspension condition were analyzed by flow cytometry following Annexin V-PE/7AAD staining (72 h of suspension culture). Statistical significance was determined using a two-sided Student's t test. The data are presented as the means ± SDs of three independent experiments. ** P<0.01 *** P<0.001, ns, not significant. SERPINE1, serine protease inhibitor clade e member 1; p-, phosphorylated; shRNA, short hairpin RNA; shSE1, shRNA targeting SERPINE1; shc, shRNA scrambled control; PLAUR, plasminogen activator, urokinase receptor; HSP90AA1, heat shock protein 90 alpha family class a member 1; RNA-seq, RNA sequencing; vec, empty vector control; siRNA, short interfering RNA; siNC, negative control siRNA; uPAR, urokinase-type plasminogen activator receptor; HSP90α, heat shock protein 90-alpha.

Article Snippet: Subsequently, suspended cells were harvested, and anoikis was quantified using an Annexin V-APC/DAPI apoptosis detection kit (cat. no. E-CK-A258; Elabscience Bionovation Inc.) according to the manufacturer's instructions.

Techniques: Western Blot, Knockdown, Control, Expressing, RNA Sequencing, Infection, Plasmid Preparation, Transfection, Activity Assay, CCK-8 Assay, Cell Culture, Suspension, Flow Cytometry, Staining, Protease Inhibitor, shRNA, Small Interfering RNA, Negative Control

Journal: iScience

Article Title: Targeting the PPAR-γ/OPTN axis to inhibit apoptosis in steroid-induced glaucoma

doi: 10.1016/j.isci.2026.115247

Figure Lengend Snippet: TMC apoptosis is observed in SIG rats (A) IOP of SIG and control rats ( n = 4, n represents the number of mice). (B) Histological analysis of HE-stained tissues. Scale bars, 20 μm. (C) The expression of Bax, Bcl-2, and Caspase-3 in the trabecular meshwork tissues was determined by IHC staining. Scale bars, 20 μm. (D) The apoptosis of trabecular meshwork tissues was determined by Tunel staining ( n = 4, n represents the number of mice). Scale bars, 100 μm. (E) The identification of TMCs by neuron-specific enolase (NSE) and fibronectin (FN). Scale bars, 50 μm. (F) Apoptosis of TMCs after dexamethasone treatment was determined by flow cytometric analysis ( n = 4, n represents the number of mice). (G) Analysis of the data in (F). (H) The apoptosis of TMCs was determined by Tunel staining ( n = 4, n represents the number of mice). Scale bars, 50 μm. (I) The expression of Bax, Bcl-2, Bcl-2/Bax, and Caspase-3 in TMCs was determined by WB staining. (J) Analysis of the data in (I) ( n = 3, n represents biological replicates). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Data in A, D, G, and J are presented as the mean ± SEM, two-tailed Student’s t test.

Article Snippet: Annexin V-APC/PI Apoptosis Kit , CatElabscience , Cat#E-CK-A217.

Techniques: Control, Staining, Expressing, Immunohistochemistry, TUNEL Assay, Two Tailed Test

Journal: iScience

Article Title: Targeting the PPAR-γ/OPTN axis to inhibit apoptosis in steroid-induced glaucoma

doi: 10.1016/j.isci.2026.115247

Figure Lengend Snippet: The apoptosis-related pathway was activated in TMCs after dexamethasone stimulation (A) The heatmap displays the DEGs between TMCs with and without dexamethasone stimulation. (B) The volcano displays the DEGs between TMCs with and without dexamethasone stimulation. (C) KEGG analysis reveals enriched pathways associated with DEGs between TMCs with and without dexamethasone stimulation. (D) The heatmap displays the DEPs between TMCs with and without dexamethasone therapy. (E) The volcano displays the DEPs between TMC with and without dexamethasone therapy. (F) KEGG analysis reveals enriched pathways associated with DEPs between TMCs with and without dexamethasone stimulation.

Article Snippet: Annexin V-APC/PI Apoptosis Kit , CatElabscience , Cat#E-CK-A217.

Techniques:

Journal: iScience

Article Title: Targeting the PPAR-γ/OPTN axis to inhibit apoptosis in steroid-induced glaucoma

doi: 10.1016/j.isci.2026.115247

Figure Lengend Snippet: Activation of PPAR-γ and Optn alleviated SIG in rats (A) Apoptosis of TMCs was determined by flow cytometric analysis ( n = 4, n represents the number of mice). (B) Analysis of the data in (A). (C) The apoptosis of TMCs was determined by Tunel staining ( n = 4, n represents the number of mice). Scale bars, 100 μm. (D) The expression of PPAR-γ, Bcl-2, Bax, Bcl-2/Bax, Caspase-3, and p-NF-κB in the TMCs was determined by WB staining. (E) Analysis of the data in (D) ( n = 3, n represents biological replicates). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Data in B, C, and E are presented as the mean ± SEM, two-tailed Student’s t test. One-way ANOVA with Tukey’s test.

Article Snippet: Annexin V-APC/PI Apoptosis Kit , CatElabscience , Cat#E-CK-A217.

Techniques: Activation Assay, TUNEL Assay, Staining, Expressing, Two Tailed Test

Journal: iScience

Article Title: Targeting the PPAR-γ/OPTN axis to inhibit apoptosis in steroid-induced glaucoma

doi: 10.1016/j.isci.2026.115247

Figure Lengend Snippet: PPAR-γ inhibited TMCs apoptosis via regulating Optn as a transcriptional factor (A) The expression of Optn in TMCs was determined by WB staining ( n = 3, n represents biological replicates). (B and C) For the dual-luciferase test, a full-length and three potential enhancer segments of the rat promoter region of Optn were created. (D) Luciferase activity was seen in the indicated groups, suggesting that PPAR-γ functioned as a transcriptional enhancer by bidding at the Optn promoter’s 3′–5′ strand’s −1925 to −1898 locations ( n = 3, n represents biological replicates). (E) A UV transilluminator was used to view the PCR products of the TMCs from the ChIP test, and PCR was used to assess the immunoprecipitated DNA ( n = 3, n represents biological replicates). ∗ p < 0.05, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001; ns, not significant. Data in A, D, and E are presented as the mean ± SEM, one-way ANOVA with Tukey’s test and two-way ANOVA with post-hoc tests.

Article Snippet: Annexin V-APC/PI Apoptosis Kit , CatElabscience , Cat#E-CK-A217.

Techniques: Expressing, Staining, Luciferase, Activity Assay, Immunoprecipitation

Journal: iScience

Article Title: Targeting the PPAR-γ/OPTN axis to inhibit apoptosis in steroid-induced glaucoma

doi: 10.1016/j.isci.2026.115247

Figure Lengend Snippet: Knockdown of the Optn gene reversed the inhibitory effect of pioglitazone on TMCs apoptosis (A) Apoptosis of TMCs was determined by flow cytometric analysis. (B) Analysis of the data in (A) ( n = 4, n represents the number of cells). (C) The apoptosis of TMCs was determined by Tunel staining. Scale bars, 100 μm. (D) The expression of Bcl-2, Bax, Bcl-2/Bax, Caspase-3, and p-NF-κB in TMCs was determined by WB staining ( n = 3, n represents biological replicates). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001; ns, not significant. Data in B and D are presented as the mean ± SEM, two-way ANOVA with post-hoc tests.

Article Snippet: Annexin V-APC/PI Apoptosis Kit , CatElabscience , Cat#E-CK-A217.

Techniques: Knockdown, TUNEL Assay, Staining, Expressing

Journal: iScience

Article Title: Targeting the PPAR-γ/OPTN axis to inhibit apoptosis in steroid-induced glaucoma

doi: 10.1016/j.isci.2026.115247

Figure Lengend Snippet: In Vivo validation: PPAR-γ/OPTN pathway protects against SIG (A) IOP of SIG, SIG+Piog, and control rats ( n = 4, n represents the number of mice). (B) Histological analysis of HE-stained tissues. Scale bars, 20 μm. (C) The apoptosis of trabecular meshwork tissues was determined by Tunel staining. Scale bars, 100 μm. (D) Analysis of the data in (C) ( n = 4, n represents the number of mice). (E) The expression of PPAR-γ, OPTN, Bcl-2, Bax, Caspase-3, and p-NF-κB in the trabecular meshwork tissues was determined by IHC staining. Scale bars, 20 μm. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Data in A and D are presented as the mean ± SEM, one-way ANOVA with Tukey’s test.

Article Snippet: Annexin V-APC/PI Apoptosis Kit , CatElabscience , Cat#E-CK-A217.

Techniques: In Vivo, Biomarker Discovery, Control, Staining, TUNEL Assay, Expressing, Immunohistochemistry

Journal: iScience

Article Title: Targeting the PPAR-γ/OPTN axis to inhibit apoptosis in steroid-induced glaucoma

doi: 10.1016/j.isci.2026.115247

Figure Lengend Snippet: A schematic model proposes the targeting of the PPAR-γ/OPTN axis, a key mechanism regulating TMCS apoptosis, as a potential strategy for intervening in SIG

Article Snippet: Annexin V-APC/PI Apoptosis Kit , CatElabscience , Cat#E-CK-A217.

Techniques: