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 Restores Mitochondrial Homeostasis and Improves Energy Metabolism BEAS-2B cells were pretreated with Res, nEVs, PD-L1@mEVs, PD-L1@nmEVs, or Res-PD-L1@nmEVs followed by H/R stimulation for subsequent analysis. (A) Representative immunofluorescence images showing the expression and localization of PINK1 (green) and the mitochondrial marker TOMM20 (red), indicating activation of mitophagy. Nuclei were stained with DAPI (blue). Scale bar: 50 μm. (B) Quantitative analysis of PINK1 fluorescence intensity. (C) Expression and localization of autophagy-related proteins LC3B and Beclin-1 detected by immunofluorescence. (D-E) Quantitative analysis of LC3B (D) and Beclin-1 (E) fluorescence intensity. (F) Mitochondrial membrane potential assessed by JC-1 staining and flow cytometry. (G) Oxygen consumption rate (OCR) profiles of lung epithelial cells under different treatments. (H-K) Key mitochondrial respiration parameters: basal respiration (H), maximal respiration (I), proton leak (J), and ATP production (K). (L) Representative confocal microscopy images of mitochondria stained with MitoTracker (green) and lysosomes stained with LysoTracker (red), demonstrating mitochondrial-lysosomal colocalization. Scale bar: 5 μm ∗ vs. Control; # vs. H/R; & vs. H/R + PD-L1@nmEVs, p < 0.05.

Article Snippet: Changes in mitochondrial membrane potential were assessed by JC-1 staining (C2003S, Beyotime).

Techniques: Immunofluorescence, Expressing, Marker, Activation Assay, Staining, Fluorescence, Membrane, Flow Cytometry, Confocal Microscopy, Control

Journal: Genes & Diseases

Article Title: PDK1 elevation was induced by epigenetic modifications of KDM3A and METTL16 to mediate TKI resistance and cancer development

doi: 10.1016/j.gendis.2025.101947

Figure Lengend Snippet: PDK1 inhibitor JX06 and gefitinib synergistically induced cell apoptosis in gefitinib-resistant lung cancer cells. (A) The protein expression levels of PDK were reduced upon the treatment of PDK1 inhibitor JX06 in PC-9 and PC-9/G cells. (B) B2B and PC-9/G cells were treated with different concentrations of JX06 for 48 h. The cell viabilities were determined by CCK-8. (C – E) The synergy effect between JX06 and gefitinib was determined and analyzed with CompuSyn software. (F) The apoptosis rates were analyzed with flow cytometry after the combined treatment of gefitinib and JX06. (G) The TUNEL assay was performed with the indicated treatment in PC-9/G cells. (H) The cells treated as described were stained with the JC-1 probe and detected using a fluorescence microscope. Red fluorescence indicates the aggregation form of JC-1, showing increased mitochondrial membrane potential (ΔΨm). Green fluorescence indicates the monomeric form of JC-1, which indicates reduced mitochondrial membrane potential (ΔΨm). Data were statistically analyzed with Student’s t -test, and values were shown as mean ± standard deviation. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: After different treatments for 48 h, the mitochondrial membrane potential of cells was detected with the enhanced mitochondrial membrane potential assay kit using JC-1 probe (Beyotime, Jiangsu, China), and the fluorescence was analyzed with a fluorescence microscope.

Techniques: Expressing, CCK-8 Assay, Software, Flow Cytometry, TUNEL Assay, Staining, Fluorescence, Microscopy, Membrane, Standard Deviation

Journal: Genes & Diseases

Article Title: PDK1 elevation was induced by epigenetic modifications of KDM3A and METTL16 to mediate TKI resistance and cancer development

doi: 10.1016/j.gendis.2025.101947

Figure Lengend Snippet: PDK1 inhibitor JX06 and gefitinib synergistically induced cell apoptosis in gefitinib-resistant lung cancer cells. (A) The protein expression levels of PDK were reduced upon the treatment of PDK1 inhibitor JX06 in PC-9 and PC-9/G cells. (B) B2B and PC-9/G cells were treated with different concentrations of JX06 for 48 h. The cell viabilities were determined by CCK-8. (C – E) The synergy effect between JX06 and gefitinib was determined and analyzed with CompuSyn software. (F) The apoptosis rates were analyzed with flow cytometry after the combined treatment of gefitinib and JX06. (G) The TUNEL assay was performed with the indicated treatment in PC-9/G cells. (H) The cells treated as described were stained with the JC-1 probe and detected using a fluorescence microscope. Red fluorescence indicates the aggregation form of JC-1, showing increased mitochondrial membrane potential (ΔΨm). Green fluorescence indicates the monomeric form of JC-1, which indicates reduced mitochondrial membrane potential (ΔΨm). Data were statistically analyzed with Student’s t -test, and values were shown as mean ± standard deviation. ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: After different treatments for 48 h, the mitochondrial membrane potential of cells was detected with the enhanced mitochondrial membrane potential assay kit using JC-1 probe (Beyotime, Jiangsu, China), and the fluorescence was analyzed with a fluorescence microscope.

Techniques: Expressing, CCK-8 Assay, Software, Flow Cytometry, TUNEL Assay, Staining, Fluorescence, Microscopy, Membrane, Standard Deviation

Journal: Materials Today Bio

Article Title: ROS-responsive hydrogels functionalized with Cu/Zn MOF targeting oxidative stress mitigation and inflammation modulation to promote spinal cord injury repair

doi: 10.1016/j.mtbio.2026.103106

Figure Lengend Snippet: Mitochondria function protection of Cu/Zn MOF@gel. (A) Representative immunofluorescence images of cells stained with MitoSOX™ Red following treatment with H 2 O 2 , Blank gel, Cu/Zn MOF, and Cu/Zn MOF@gel. Scale bar = 50 μm. (B) Evaluation of mitochondrial membrane potential (ΔΨm) using JC-1 staining in PC12 cells by flow cytometry. (C) Representative flow cytometry histograms of intracellular calcium ion (Ca 2+ ) levels in PC12 cells using Fluo-4 AM staining. (D) Quantitative analysis of the mean fluorescence intensity of MitoROS. (E – F) Quantitative statistical analysis of the percentage of JC-1 monomers (E) and JC-1 aggregates (F) . (G) Quantitative statistical analysis of the percentage of Fluo-4 positive cells. (H) Representative transmission electron micrographs of mitochondria in PC12 cells. Data are presented as mean ± SD (n = 3). One-way ANOVA with Tukey's post-hoc test was employed. Statistical significance is indicated as ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001. “ns” means non-significant results. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Dichlorodihydrofluorescein diacetate kit (DCFH-DA), Dihydroethidium kit (DHE), CCK-8 Kit, Calcein/PI cell viability/cytotoxicity assay kit, Nitroblue tetrazolium (NBT), Enhanced mitochondrial membrane potential assay kit with JC-1, Mitochondrial Superoxide Assay Kit with MitoSOTM Red, and Luxol fast blue (LFB) were purchased from Shanghai Beyotime Biotechnology Co., Ltd. Dulbecco's modified eagle medium (DMEM), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12), Poly-D-lysine (PDL), Neurobasal-A medium (NB), B-27 Serum-Free Supplement (B-27), Hank's balanced salt solution (HBSS), Trypsin, fetal bovine serum (FBS) and phosphate buffer solution (PBS) were obtained from Gibco.

Techniques: Immunofluorescence, Staining, Membrane, Flow Cytometry, Fluorescence, Transmission Assay

Journal: Materials Today Bio

Article Title: Dual-targeted manganese-doped carbon dots activate the cGAS–STING pathway and immunogenic cell death for potent glioblastoma immunotherapy

doi: 10.1016/j.mtbio.2026.103073

Figure Lengend Snippet: Mn-CDs induce mitochondrial dysfunction and immunogenic cell death (ICD) via robust ROS generation. Intracellular ROS generation in CT2A cells treated with PBS, CDs, or Mn-CDs for 4 h. (A) Representative confocal images using DCFH-DA staining (green fluorescence). Scale bar: 50 μm. (B) Flow cytometric analysis and (C) corresponding quantitative analysis of ROS-positive cell populations (n = 3). The results indicate a surge in ROS levels in the Mn-CDs group. (D) UV–vis absorption spectra of Methylene Blue (MB) mixed with H 2 O 2 and different concentrations of Mn-CDs. The degradation of MB indicates the generation of hydroxyl radicals (·OH) via manganese-mediated Fenton-like reactions. Analysis of mitochondrial membrane potential using JC-1 staining. (G) Confocal images of CT2A cells treated with PBS, CDs (25, 50 μg/mL), or Mn-CDs (25, 50 μg/mL). Red (aggregates) and green (monomers) fluorescence represent high and low mitochondrial membrane potential, respectively. Scale bar: 100 μm. (E) Quantitative analysis of the red/green fluorescence percentage. (F) Quantification of extracellular ATP release in the culture supernatant of CT2A cells. (J) Representative confocal images and (H) quantitative analysis of CT2A cells nuclear fluorescence intensity of HMGB1. The decrease in nuclear intensity indicates the release of HMGB1 from the nucleus. Scale bar: 100 μm. (K) Representative confocal images and (I) quantitative analysis of CRT exposure on the surface of CT2A cells. Scale bar: 100 μm. Transmission electron microscopy (TEM) images showing mitochondrial ultrastructure in (L) CT2A and (M) GL261 cells treated with PBS, CDs, or Mn-CDs. Scale bars: 2 μm (low magnification) and 500 nm (high magnification inset). Mn-CDs treatment induces mitochondrial swelling and cristae disruption. Data are presented as mean ± SD (∗∗∗∗p < 0.0001; ns: no significance).

Article Snippet: The JC-1 Assay Kit, DAPI, Mito-Tracker Green, Crystal Violet Staining Solution, and Protease/Phosphatase Inhibitor Cocktail (P1045) were purchased from Beyotime Biotechnology (Shanghai, China).

Techniques: Staining, Fluorescence, Membrane, Transmission Assay, Electron Microscopy, Disruption