annexin v fitc apoptosis detection kit  (Elabscience Biotechnology)

Journal: Advanced Science

Article Title: Phosphorylated STYK1 Promotes Gemcitabine Resistance via Autophagy‐Mediated Cancer Stemness in Pancreatic Cancer

doi: 10.1002/advs.202521488

Figure Lengend Snippet: STYK1 promotes gemcitabine resistance in pancreatic cancer cells. (A, B) PANC‐1 cells were treated with different concentrations of gemcitabine for the indicated time periods, and STYK1 mRNA expression was assessed by RT‐qPCR (n = 6). **** p < 0.0001 according to Student's t ‐test. (C) Western blotting analysis of STYK1 expression levels in human pancreatic cancer cell lines PANC‐1 and AsPC‐1, as well as in the gemcitabine‐resistant cell lines PANC‐1/GEM and AsPC‐1/GEM. D) RT‐PCR analysis of STYK1 mRNA levels in PANC‐1, AsPC‐1, PANC‐1/GEM and AsPC‐1/GEM cells. **** p < 0.0001 according to Student's t ‐test. (E) Representative images of IHC staining for STYK1 in gemcitabine‐sensitive and resistant pancreatic cancer PDX tumors tissues. (F–I) Cell viability of PANC‐1/GEM and AsPC‐1/GEM cells with stable STYK1 knockdown, as well as PANC‐1 and AsPC‐1 cells stably overexpressing STYK1, following treatment with different concentrations of gemcitabine for 72 h, as determined by CCK‐8 assay. (J) PANC‐1/GEM cells with stable STYK1 knockdown were treated with gemcitabine (5 µM) for different durations as indicated, and OD values were assessed using the CCK‐8 assay. **** p < 0.0001 according to Student's t ‐test. (K–M) Apoptosis of PANC‐1/GEM and AsPC‐1/GEM cells with stable STYK1 knockdown following gemcitabine treatment (5 µM for PANC‐1/GEM and 40 µM for AsPC‐1/GEM, 24 h), assessed using an Annexin V‐AbFluor 488/PI double staining Apoptosis Detection Kit. Annexin V‐positive cells indicate early apoptosis, whereas PI‐positive cells indicate late apoptosis/necrosis (n = 3). **** p < 0.0001 according to Student's t ‐test. (N) Representative images of resected tumors from nude mice in different groups (n = 5). O) Tumor weights of resected tumors in different groups of mice (n = 5). **** p < 0.0001 according to Student's t ‐test. ** p < 0.01; **** p < 0.0001 according to Student's t ‐test. (P) Tumor volume change curves in different groups of mice (n = 5); Tumor volume was measured every 5 days after cell injection and during gemcitabine treatment (25 mg/kg). **** p < 0.0001 according to Student's t ‐test. (Q) Representative H&E staining and Ki67 immunohistochemical staining images of resected tumor tissues (n = 5). Scale bar: 50 µm. Data were expressed as mean ± SD, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 according to Student's t ‐test.

Article Snippet: Apoptosis was assessed using an Annexin V‐FITC Apoptosis Detection Kit (Elabscience, E‐CK‐A252).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Analysis, Reverse Transcription Polymerase Chain Reaction, Immunohistochemistry, Knockdown, Stable Transfection, CCK-8 Assay, Double Staining, Injection, Staining, Immunohistochemical staining

Journal: Cancer Medicine

Article Title: Pan‐Cancer Analysis of the Immunomodulatory Roles and Prognostic Value of PDXP and Experimental Verification in SKCM

doi: 10.1002/cam4.72311

Figure Lengend Snippet: Association of PDXP expression with multiple signaling pathways and biological functions. (A) PDXP expression was significantly positively correlated with DNA Repair, G2M Checkpoint, and E2F Targets in most cancer types. (B) PDXP expression was highly enriched in the Cell Cycle pathway across most cancer types. In SKCM, PDXP expression was potentially associated with the activation of cell cycle and TSC/mTOR pathways, while inversely associating with the Apoptosis pathway. (C) A total of 402 genes significantly correlated with PDXP expression in SKCM were selected for enrichment analysis. (D) KEGG pathway analysis revealed potential enrichment in the mTOR signaling pathway, lysosome, and cell cycle pathways. (E) GO functional enrichment analysis showed significant enrichment in mitochondrial function and protein metabolic processes. *** p < 0.001.

Article Snippet: For detection of cell apoptosis, 5 × 10 5 A2058 and A375 cells were resuspended in 500 μL of diluted 1 × Annexin V Binding Buffer from the Annexin V‐APC/DAPI Apoptosis Kit (cat. no. E‐CK‐A258; Elabscience).

Techniques: Expressing, Protein-Protein interactions, Activation Assay, Analysis, Functional Assay

Journal: Cancer Medicine

Article Title: Pan‐Cancer Analysis of the Immunomodulatory Roles and Prognostic Value of PDXP and Experimental Verification in SKCM

doi: 10.1002/cam4.72311

Figure Lengend Snippet: PDXP promotes melanoma cell proliferation and suppresses apoptosis potentially via the AKT/mTOR signaling pathway. (A) Flow cytometry analysis showed abnormal accumulation of cells in the G2/M phase in the shPDXP#3 group. (B) The proportions of early apoptotic (Q3) and late apoptotic (Q2) cells increased significantly in the shPDXP#3 group. (C) Western blot results showed that PDXP knockdown downregulated the expression of Bcl‐2 and Bcl‐xL, and upregulated the expression of Bax. (D) PDXP knockdown significantly inhibited the phosphorylation of AKT and mTOR and increased PTEN expression. ** p < 0.01, *** p < 0.001.

Article Snippet: For detection of cell apoptosis, 5 × 10 5 A2058 and A375 cells were resuspended in 500 μL of diluted 1 × Annexin V Binding Buffer from the Annexin V‐APC/DAPI Apoptosis Kit (cat. no. E‐CK‐A258; Elabscience).

Techniques: Flow Cytometry, Analysis, Western Blot, Knockdown, Expressing, Phospho-proteomics

Journal: Blood Cancer Journal

Article Title: The role of RAD23A in DNA repair and proteasome-dependent protein degradation in multiple myeloma

doi: 10.1038/s41408-026-01574-z

Figure Lengend Snippet: A Proteomic-based GSEA Reactome showing negative enrichment of the deubiquitination pathway following RAD23A knockout (NES = −1.398, FDR q -value = 0.104). B MM.1R and AMO1 RAD23A-iKO cells, as well as H929, ANBL6-WT, and ANBL6-BR RAD23A-iKD cells, were cultured with Dox as described in Fig. . Protein lysates were subjected to immunoblotting using antibodies against K48-linked polyubiquitin and β-actin. CT = control. C Aggresome and K48-linked polyubiquitinated protein (K48-polyU) accumulation were detected using an aggresome detection kit and anti-K48-polyubiquitin antibody, followed by confocal microscopy (60×). D MM.1R RAD23A-iKO cells were cultured with Dox for 9 days. MM.1R cells treated with DMSO or 1 µM bortezomib for 3 h were used as controls. Cell pellets were harvested and lysed. 20S proteasome activities (CTL, chymotrypsin-like; TL, trypsin-like; CL, caspase-like) were measured. Proteasome activity was normalized to the DMSO control or parental MM.1R control cells (mean ± SD, n = 3, assessed by two-way ANOVA with Tukey’s test). E MM.1R and AMO1 RAD23A-iKO cells, as well as H929, ANBL6-WT and ANBL6-BR RAD23A-iKD cells, were prepared as described in Fig. . Cells were lysed and immunoblotted using antibodies against p-eIF2α, ATF4, CHOP, PARP, cleaved caspase-3, and β-actin. F Caspase-3/7 activation was detected using the IncuCyte Caspase-3/7 Green Dye and live-cell imaging. G Apoptosis was analyzed by flow cytometry using Annexin V-FITC/DAPI double staining (mean ± SD, n = 3, assessed by two-way ANOVA with Sidak’s test).

Article Snippet: Apoptosis was further validated by flow cytometric analysis using an Annexin V-FITC/DAPI apoptosis detection kit (Elabscience, #E-CK-A252), following the manufacturer’s protocol.

Techniques: Knock-Out, Cell Culture, Western Blot, Control, Confocal, Microscopy, Activity Assay, Activation Assay, Live Cell Imaging, Flow Cytometry, Double Staining