annexin v apc pi apoptosis detection kit  (Elabscience Biotechnology)

Journal: Cell Reports Medicine

Article Title: CAR-M2 immunotherapy resolves renal fibrosis via revascularization and apoptosis of profibrotic Cxcr2 + endothelial cells

doi: 10.1016/j.xcrm.2026.102698

Figure Lengend Snippet: The secretion of MMP2 by CAR-M2 activates the transcription factor Rxra in Cxcr2 + ECs (A) Position relationship between CXCR2 + ECs and FAP + fibroblasts was visualized by immunofluorescence staining. Blue: DAPI, green: CD31, red: CXCR2, French gray: FAP. (B) Volcano plot of differentially expressed genes before and after co-culture of CAR-M2 with FAP + fibroblasts. (C) Representative Masson and Sirius red staining images of mice kidney tissue sections following the injection of PBS into the renal subcapsule (Sham and UUO), CAR-M2-loaded HAMA-CS hydrogel injected into the renal subcapsule (CAR-M2), and PBS into the renal subcapsule of MMP2 inhibitor mice (MMP2 inhibitor). Detection at day 7. Scale bars, 100 μm. (D) Average collagen deposition density by Masson staining in (C), n = 5. (E) Average collagen deposition density (the percentage of positively stained area per HPF) by Sirius red staining in (C), n = 5. (F) Re-clustering of endothelial cells from human kidney single-cell sequencing data based on regulon activity, t-SNE (t-distributed stochastic neighbor embedding) displayed by cell type (left), and kidney condition (right). (G) SCENIC analysis of cell stability in clusters derived from regulon activity-based dimensionality reduction. (H) Ranked plot of transcription factor activity in CXCR2 + ECs. (I) Differential expression of the Rxra gene among groups in mouse single-cell transcriptome samples. (J) Changes in the endothelial transcription factor RXRA after CAR-M2 treatment and in MMP2 inhibitor-treated mice. Blue: DAPI, red: CD31, green: Rxra, yellow indicates the co-labeling of CD31 with Rxra, Scale bars, 25 μm. Arrowheads indicate the double positive cells. (K) Statistical analysis of the number of CD31 + Rxra + cells in (J), n = 5. (L) Cell co-culture protocol to verify that CAR-M2 activates the transcription factor Rxra by secreted MMP2. (M) After cell co-culture using protocol in (L), the lentiviral constructs Cxcr2 + ECs were collected to detect the changes in their Rxra and Cxcr2 expression levels. (N) Average collagen deposition density by Masson staining, mice kidney tissue sections following the injection of PBS into the renal subcapsule (Control), CAR-M2-loaded HAMA-CS hydrogel injected into the renal subcapsule (CAR-M2), and PBS into the renal subcapsule of Rxra agonist and inhibitor mice (Rxra agonist and Rxra inhibitor). Detection at day 14. n = 5. (O) Average collagen deposition density by Sirius red staining, mice kidney tissue sections following the injection of PBS into the renal subcapsule (Control), CAR-M2-loaded HAMA-CS hydrogel injected into the renal subcapsule (CAR-M2), and PBS into the renal subcapsule of Rxra agonist and inhibitor mice (Rxra agonist and Rxra inhibitor). Detection at day 14. n = 5. (P) Mitochondrial autophagy-related gene score in human normal and CKD groups. (Q) Apoptotic-related gene score in human normal and CKD groups. (R) Mitochondrial autophagy-related gene score in mice Sham, UUO, and CAR-M2 groups. (S) Apoptotic-related gene score in mice Sham, UUO, and CAR-M2 groups. (T) The changes in the expression levels of autophagy-related protein PINK1 and apoptosis-related protein active caspase-3. (U) The cell apoptosis rate after the aforementioned (T) co-culture was detected by flow cytometry, and the proportion of Annexin-APC positive cells was counted as the cell apoptosis rate. The detection time was 6 h after co-culture. n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns, not significant (one-way ANOVA). See also .

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

Techniques: Immunofluorescence, Staining, Co-Culture Assay, Injection, Single Cell, Sequencing, Activity Assay, Derivative Assay, Quantitative Proteomics, Labeling, Construct, Expressing, Control, Flow Cytometry

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:

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