annexin v apc apoptosis detection kit  (Multi Sciences (Lianke) Biotech Co Ltd)

Journal: Cellular Oncology

Article Title: BCAA catabolism mediates POU2AF1 propionylation to enhance T-ALL development

doi: 10.1007/s13402-026-01201-w

Figure Lengend Snippet: BCAT1 regulates T-ALL cell fate decisions. ( A ) Representative flow cytometry plots of leukemia cells (GFP + ) in the peripheral blood of recipient mice 3 weeks after secondary transplantation. ( B ) Quantification of leukemia cells (GFP + ) in peripheral blood after secondary transplantation ( n = 6). ( C ) Representative images of the size of the liver, spleen and thymus from recipient mice post-secondary transplantation. ( D ) Quantification of the data shown in ( C ) ( n = 6). ( E ) Overall survival was determined for the leukemic mice shown in ( B ) ( n = 6). ( F ) Representative flow cytometry plots of leukemia cells (GFP + ) in the peripheral blood of recipient mice 3 weeks after tertiary transplantation. ( G ) Quantification of leukemia cells (GFP + ) in peripheral blood after tertiary transplantation ( n = 6). ( H ) Organ weight quantification of the liver, spleen and thymus from transplanted mice ( n = 4). ( I ) Overall survival was determined for the leukemic mice shown in ( G ) ( n = 10). ( J ) Representative flow cytometry plots showing early and late apoptosis of WT and KO T-ALL cells in peripheral blood after secondary transplantation. ( K ) Quantification of apoptotic populations from Panel ( J ) ( n = 3). ( L ) Flow cytometric analysis of the homing efficiency of WT and KO T-ALL cells to bone marrow 16 h post-injection. ( M ) Quantification of homing efficiency, shown in ( L ) ( n = 5). ( N ) Cell cycle status was determined in WT and KO T-ALL cells of the recipients. ( O )Quantification data of the phases of the cell cycle in Panel ( N ) ( n = 3). The data are presented as the means ± SDs. Student’s two-tailed unpaired t test (B, G, M) , log-rank test ( E , I ) and two-way ANOVA with Sidak’s multiple comparison test ( D , H , K , O ) were used for the comparison of statistical significance (*, P < 0.05; **, P < 0.01; and ***, P < 0.001)

Article Snippet: Apoptosis was assessed using an Annexin V-APC Apoptosis Detection Kit (MULTI SCIENCE, AP105) according to the manufacturer’s protocol.

Techniques: Flow Cytometry, Transplantation Assay, Injection, Two Tailed Test, Comparison

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: Frontiers in Oncology

Article Title: ITPKA suppresses glioma progression and predicts patient prognosis

doi: 10.3389/fonc.2026.1802857

Figure Lengend Snippet: Effect of ITPKA overexpression on the functional characteristics of GBM cells. Flow cytometry analysis (A, B) shows the effect of upregulated ITPKA expression on the cell cycle progression of U251-MG and T98G cells; (C, D) evaluates the effect of upregulated ITPKA expression on the apoptosis level of the two indicated cell lines. All experimental data are presented as mean ± standard deviation (SD) from six independent repeated experiments, and Student’s t-test was used for statistical analysis. Data annotation: ns: not significant; *p < 0.05, ****p < 0.0001.

Article Snippet: A total of 5×10 5 stably transfected U251-MG and T98G cells were centrifuged, after which apoptosis detection was performed using the Annexin V-APC/PI Double Staining Apoptosis Detection Kit (KGA1107-100; Keygen Biotech, Nanjing, China).

Techniques: Over Expression, Functional Assay, Flow Cytometry, Expressing, Standard Deviation

Journal: Frontiers in Oncology

Article Title: ITPKA suppresses glioma progression and predicts patient prognosis

doi: 10.3389/fonc.2026.1802857

Figure Lengend Snippet: Effect of ITPKA knockdown on the functional characteristics of GBM cells. Flow cytometry analysis (A, B) shows the effect of downregulated ITPKA expression on the cell cycle progression of U251-MG and T98G cells; (C, D) evaluates the effect of downregulated ITPKA expression on the apoptosis level of the two indicated cell lines. All experimental data are presented as mean ± standard deviation (SD) from six independent repeated experiments, and Student’s t-test was used for statistical analysis. Data annotation: ns: not significant; *p < 0.05, **p < 0.01, ****p < 0.0001.

Article Snippet: A total of 5×10 5 stably transfected U251-MG and T98G cells were centrifuged, after which apoptosis detection was performed using the Annexin V-APC/PI Double Staining Apoptosis Detection Kit (KGA1107-100; Keygen Biotech, Nanjing, China).

Techniques: Knockdown, Functional Assay, Flow Cytometry, Expressing, Standard Deviation

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