annexin v apc  (Keygen Biotech)

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: bioRxiv

Article Title: Intranuclear polyglycine aggregation drives neurodegeneration through epigenetic repression of chromatin accessibility and transcription

doi: 10.64898/2026.05.11.723935

Figure Lengend Snippet: (A) Schematic of the polyG constructs used in this study. Gly7 served as a short-repeat control, whereas Gly73 contained an expanded 73-glycine tract. To bias the subcellular distribution of aggregates, Gly73 was fused to either a 2×NLS or an NES, generating Gly73-NLS and Gly73-NES, respectively. All constructs were expressed under the CAG promoter and fused to 2×EGFP and a 3×FLAG tag. (B) Representative confocal images of SH-SY5Y cells expressing the indicated constructs. Gly73-NLS predominantly formed intranuclear aggregates, whereas Gly73-NES mainly formed extranuclear aggregates, with Gly73 showing an intermediate distribution. Boxed regions are shown at higher magnification. Scale bar, 50 μm. (C) Quantification of the percentage of intranuclear aggregates among total aggregates in cells expressing the indicated constructs (n=5). (D) LDH release assay showing cytotoxicity induced by the indicated constructs in SH-SY5Y cells (n=11). (E) Quantification of Annexin V-APC-positive cells by flow cytometry, showing increased apoptosis in the Gly73-NLS group (n=3). (F) Representative flow cytometry gating plots used for Annexin V-APC analysis. (G) Representative histograms of Annexin V-APC fluorescence intensity in cells expressing the indicated constructs. Data are means ± SEM and analyzed with one-way ANOVA ( C-E ). ‘‘ns’’ represents non-significant; *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: The cell pellets were washed with Hank’s Balanced Salt Solution (HBSS), gently resuspended, and incubated with Annexin V-APC (KGA1105-20, KeyGEN BioTECH) for 10 min in the dark according to the manufacturer’s instructions.

Techniques: Construct, Control, Expressing, Lactate Dehydrogenase Assay, Flow Cytometry, Fluorescence