Journal: Molecular medicine reports

Article Title: BCG stimulation promotes dendritic cell proliferation and expression of VDR and CYP27B1 in vitamin D‑deficient mice.

doi: 10.3892/mmr.2019.10780

Figure Lengend Snippet: Figure 2. Effect of BCG on BMDC differentiation and maturation. (A) Cell morphology during BMDC induction from days 1 to 6, as observed by light micros- copy. (B) BCG stimulated BMDC maturation. (C) Phenotypic changes of the BMDCs induced by BCG stimulation and (D) the levels of the surface molecules of DCs. All images were captured under identical magnification and microscopy conditions. **P<0.01 vs. 0 mg/ml in the normal control or vitamin D‑deficiency groups. ##P<0.01, vitamin D‑deficiency group vs. normal control group. Scale bar, 20 µm. BCG, Bacillus Calmette‑Guérin; BMDC, bone marrow‑derived dendritic cell; CD11c, integrin alpha‑X; CD80, T‑lymphocyte activation antigen CD80; MHC‑I, major histocompatibility complex class I; MHC‑II, major histocompatibility complex class II; CD86, T‑lymphocyte activation antigen CD86.

Article Snippet: The cell concentration was adjusted to 1x106 cells/ml and the cells were fixed with 0.1 ml polyformaldehyde for 30 min at 25 ̊C Subsequently, fluorescein isothiocyanate‐labeled integrin alpha‐X (CD11c; 1:50; 130‐110‐700; Miltenyi Biotech), T‐lymphocyte activation antigen CD86 (CD86; 1:50; 130‐102‐506; Miltenyi Biotech), T-lymphocyte activation antigen cd80 (cd80; 1:50; 130‐116‐459; Miltenyi Biotech), major histocompatibility complex class i (MHc-i; 1:50; ab95572; abcam) and MHC‐II (1:50; 130‐112‐386; Miltenyi Biotech) were added, respectively.

Techniques: Microscopy, Control, Activation Assay, Immunopeptidomics

Journal: Journal of nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages.

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: Fig. 2 LpEVs alleviated lung inflammation and injury in mice. A. Schematic depiction of the experiment. B. Representative images showing histopatho logical alterations in lungs of different treatment groups. ICR mice (n = 5) were sitimulated with LPS (2.5 mg/kg, intratracheally) for 3 h, then administered PBS or LpEVs (100 µg) via the tail vein injection method. Mice were euthanized 24 h post-treatment. Scale bar = 200 μm. C. Representative F4/80 and MPO IHC staining images in different lungs. Scale bars represent 50 μm. D. Protein concentration in BALF. E. Cell counts in BALF. F. The concentration of cytokines/chemokines in the BALF was assessed using ELISA. G. Flow cytometry analysis of neutrophils (CD11b+ Ly6G+) in the BALF. H. Flow cytometry analysis of macrophages (CD11b+ F4/80+) in the BALF. I. Flow cytometry analysis of alveolar macrophages (CD11c+ Siglec F+) in the BALF

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702- 82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: Injection, Immunohistochemistry, Protein Concentration, Concentration Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Journal of nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages.

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: Fig. 4 LpEVs treatment inhibited LPS-induced ferroptosis in vivo. A. The levels of reactive oxygen species (ROS) in lung BALF cells were evaluated using DCFH-DA. The left panel displays a flow cytometry diagram, while the right panel shows the mean fluorescence intensity of ROS. B. Relative MDA content in lungs. C. Relative Fe2+ content in lungs. D. Relative GSH content in lungs. E. Relative mRNA expression of ferroptotic-related genes were analyzed by qRT-PCR. F. The expression levels of ferroptotic-related proteins in lungs were analyzed by western blotting. ACTIN is adopted as the loading control. G. Representative IHC staining images for GPX4, SLC7A11 and ACSL4 were obtained from different lung samples. Scale bars represent 50 μm. H. The statisti cal graph of the expression of GPX4, SLC7A11 and ACSL4 in IHC staining. I. Representative IHC staining images for macrophages M1 marker CD86 and M1 marker CD206 in lungs. Scale bars represent 50 μm. J. The statistical graph of the expression of CD86 and CD206 in IHC staining. Quantification of proteins expresion levels in IHC staining were determined by image J software. The values for the LPS-PBS and LPS-EV were expressed as percentages of the values for the Sham group. *P < 0.05, **P < 0.01, ***P < 0.001, ***P < 0.0001 and ns, no significant difference

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702- 82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: In Vivo, Flow Cytometry, Fluorescence, Expressing, Quantitative RT-PCR, Western Blot, Control, Immunohistochemistry, Marker, Software

Journal: Journal of nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages.

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: Fig. 5 LpEVs facilitated the transition of macrophages towards an anti-inflammatory phenotype. A. Confocal laser scanning microscopy images dis played the presence of Dio-labeled LpEVs in MH-S cells. Green: Dio labeled LpEVs; Blue: Hoechst. Scale bar = 20 μm. B. The Dio-positive MH-S cells were determined by flow cytometry. C. Relative mRNA expression levels of inflammatory factors were assessed by qRT-PCR. M1 proinflammatory genes: Il1β, Il6, Tnfα, and iNos, M2 anti-inflammatory genes: Arg-1, and Cd206. The M1 macrophages were induced by LPS treatment, and M2 macrophages were induced by IL-4 treatment. D. The levels of pro-inflammatory and anti-inflammatory cytokines in the medium supernatants were quantified using ELISA. E. The expression of CD86 (an M1 marker) on MH-S cells 12 h after exposure to LPS was assessed using flow cytometry, with representative histograms and average relative mean fluorescence intensity (MFI) provided. F. Flow cytometry was used to detect the expression of CD206, an M2 marker, on MH-S cells 12 h after exposure to IL4

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702- 82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: Confocal Laser Scanning Microscopy, Labeling, Flow Cytometry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Marker, Fluorescence

Journal: Journal of nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages.

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: Fig. 6 LpEVs attenuated ferroptosis in macrophages. A. The ROS level in MH-S was assessed by flow cytometry. B. Relative MDA content in MH-S. C. Rela tive GSH content in MH-S. D. The intercellular Fe2+ was detected by FerroOrange (a specific probe of Fe2+). Scale bar = 100 μm. E. The expression levels of ferroptotic-related proteins and NRF2-related proteins in MH-S were detected by western blot. ACTIN is adopted as the loading control. F. SLC7A11, GPX4 and ACSL4 immunofluorescent staining results of differently treated MH-S. Scale bar = 20 μm. G. NRF2 and HO-1 immunofluorescent staining results of differently treated MH-S. Scale bar = 20 μm

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702- 82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: Flow Cytometry, Expressing, Western Blot, Control, Staining

Journal: Journal of nanobiotechnology

Article Title: Lactiplantibacillus plantarum -derived extracellular vesicles alleviate acute lung injury by inhibiting ferroptosis of macrophages.

doi: 10.1186/s12951-025-03405-y

Figure Lengend Snippet: Fig. 7 LpEVs regulated ferroptosis in macrophages via delivery of cbn-let-7. A. miRNA microarray analyze the miRNA expression profile of LpEVs. B. The 4 miRNAs (cel-mir-4937, cbn-let-7, ngi-miR-92a and gsa-mir-9394b) expression level in LpEVs. C. cbn-let-7 expression in PBS and LpEVs treated MH-S cells. D, E. Structure and luciferase result of dual luciferase reporter gene. F. The relative expression of cbn-let-7 in NC, cbn-let-7 mimic and inhibitor treated MH-S cells. G. The protein expression of of ACSL4 and GPX4 in NC, cbn-let-7 mimic and inhibitor treated MH-S cells. H. The expression of cbn-let-7 in different treated cells. I. The protein expression of of ACSL4 and GPX4 in different treated cells. ACTIN is adopted as the loading control. J. The intercellular Fe2+ in different treated cells was detected by FerroOrange (a specific probe of Fe2+)

Article Snippet: The population of alveolar macrophages (anti-CD11c, anti-Siglec F) (E-AB-F0991E, Elabscience, Wuhan, China) (12-1702- 82, Thermo Fisher, USA), macrophages (anti-F4/80, anti-CD11b) (E-AB-F0995D, E-AB-F1081E, Elabscience, Wuhan, China), and neutrophils (anti-CD11b, anti-Ly6G) (E-AB-F1081E, E-AB-F1108C, Wuhan, China) were assessed using flow cytometry with specific antibodies.

Techniques: Microarray, Expressing, Luciferase, Control

Journal: Cell Death & Disease

Article Title: Oncolytic peptide LTX-315 plus an anti-CTLA-4 antibody induces a synergistic anti-cancer immune response in residual tumors after radiofrequency ablation of hepatocellular carcinoma

doi: 10.1038/s41419-025-07622-z

Figure Lengend Snippet: A Experimental workflow for silencing the cGAS-STING pathway and ICD in vivo studies. B , C Western blotting was used to confirm the knockout of cGAS and STING proteins in Hepa1-6 cells. D , E The proportion of mature dendritic cells (CD80 + CD86 + DCs) was assessed after treatment with the anti-CD11c antibody. F , G Western blotting was applied to confirm the reduction of ICD-related molecules (ATP, CALR, HMGB1) after treatment with the anti-CD11c antibody. H , I Phosphorylation of TBK1 and IRF3 was reduced in tumors after silencing the cGAS-STING pathway. J , K Tumors were harvested and weight was assessed, showing an increase in tumor size after silencing the cGAS-STING pathway and ICD in all five groups. L – N Flow cytometry analysis was used to evaluate the levels of functional CD8 + T cells after silencing the cGAS-STING pathway and ICD in all five groups. n = 6 per group. Error bars represent standard deviation. Ns not statistically significant. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: CD11c monoclonal antibodies (BioXCell, NH, USA) were administered intraperitoneally at 100 μg/mouse, starting 7 days before combination therapy and repeated weekly for 3 weeks.

Techniques: In Vivo, Western Blot, Knock-Out, Phospho-proteomics, Flow Cytometry, Functional Assay, Standard Deviation

Journal: iScience

Article Title: Peripheral immune landscape in pancreatic ductal adenocarcinoma reveals expansion of effector states with disease progression

doi: 10.1016/j.isci.2026.115034

Figure Lengend Snippet: Machine learning classification identifies predictive immune markers distinguishing PDAC from healthy individuals (A) Schematic overview of the machine learning workflow used to identify predictive immune markers from high-dimensional spectral flow cytometry data. Peripheral blood samples from 38 healthy individuals and 39 treatment-naive PDAC patients (stages I–IV) were analyzed. Mean fluorescence intensity (MFI) values of surface markers were extracted from CD45 + cells, asinh-normalized, and aggregated at the individual level. The dataset was split into 70% training and 30% testing sets for model development and evaluation. (B–D) Top 10 predictive markers ranked by feature importance scores across three independent classifiers: random forest (RF; B), gradient boosting (GB; C), and Bayesian additive regression tree (BART; D). Markers are ranked by their Gini importance scores. (E and F) Receiver operating characteristic (ROC) curves showing model performance in classifying healthy versus PDAC samples (E) and in predicting PDAC stages (F). Curves represent random forest (orange), GB (green), and BART (blue) classifiers, with area under the curve (AUC) values indicating model accuracy. (G and H) Overall expression levels of CD95 and CD45RA, and (H) the representative expression patterns of CD95 in PD1+ CD4 + T cells, CD45RA+ terminal effector (TE) CD8 + T cell, CD11c+ dendritic cells (DCs), and non-classical monocytes (Mo) of healthy individuals and PDAC patients. Each bar represents Z score normalized mean fluorescence intensity (MFI) across individuals. The data are represented as mean ± SEM, and the t test p value, indicating statistical significance, is displayed at the top of each violin and boxplot. ∗∗∗ p < 0.001; NS, non-significance.

Article Snippet: violetFlour 450 Anti-Human CD11c (Clone: 3.9) , Cytek Biosciences , Cat# 75–0116_T100; PRID: AB_2621938.

Techniques: Flow Cytometry, Fluorescence, Biomarker Discovery, Expressing