Journal: Journal of Biomedical Research

Article Title: HDGF derived from Müller cells enhances the activation of microglia in diabetic retinopathy

doi: 10.7555/JBR.38.20240386

Figure Lengend Snippet: HDGF promoted microglia activation and IL-1β secretion. A: Analysis of a published list of target genes of HDGF integrated with mouse retinal single-cell RNA sequencing data, indicating predominant target cell types. The violin plot shows expression of the target gene Itgb2 in microglia from control and streptozotocin (STZ)-induced diabetic mouse retinas. The microglia cluster is indicated. B: Enzyme-linked immunosorbent assay measuring IL-1β secretion in the supernatant of BV2 microglial cells. Cells were treated under low-glucose (LG, 1 g/L) or high-glucose (HG, 5 g/L) conditions for 48 h, with or without recombinant HDGF (rHDGF) and an HDGF-neutralizing antibody (anti-HDGF) ( n = 3). C: Quantitative PCR analysis of Itgb2 mRNA expression in BV2 cells treated as indicated ( n = 3). D: Representative immunofluorescence images (left) and quantification (right) of ITGB2-positive BV2 cells following the treatments. Nuclei were counterstained with DAPI. Quantitative analysis of ITGB2-positive cells treated as indicated ( n = 3). E: Representative immunofluorescence images (left) and quantification (right) of CD68-positive BV2 cells following the treatments. Nuclei were counterstained with DAPI. Quantitative analysis of CD68-positive cells treated as indicated ( n = 3). F: Feature plot from retinal scRNA-seq data showing specific Itgb2 expression within the microglia cluster (outlined). G: Western blotting analysis of ITGB2 protein expression in BV2 microglial cells and primary Müller cells. α-Tubulin served as the loading control. Quantitative protein levels of ITGB2 in BV2 and Müller cells ( n = 3 per group). Data are presented as mean ± standard deviation. Statistical significance was determined by one-way analysis of variance (ANOVA; B–E) or paired Student's t -test (G). * P < 0.05; ** P < 0.01; *** P < 0.001. Abbreviations: HDGF, hepatoma-derived growth factor; ITGB2, integrin beta 2.

Article Snippet: The immortalized murine microglial cell line (BV2) (Cat. #CRL-2469) was purchased from the American Type Culture Collection and cultured in DMEM/F12 medium (Gibco) supplemented with 10% FBS (Invitrogen), 1% glutamine (Invitrogen), and streptomycin (100 U/mL; Invitrogen) at 37 °C, 5% CO 2 .

Techniques: Activation Assay, RNA Sequencing, Expressing, Control, Enzyme-linked Immunosorbent Assay, Recombinant, Real-time Polymerase Chain Reaction, Immunofluorescence, Western Blot, Standard Deviation, Derivative Assay

Journal: Journal of Biomedical Research

Article Title: HDGF derived from Müller cells enhances the activation of microglia in diabetic retinopathy

doi: 10.7555/JBR.38.20240386

Figure Lengend Snippet: High glucose upregulated the expression of HDGF receptor in microglia. A: Western blotting analysis of HDGF protein expression in BV2 microglial cells cultured under low-glucose (LG, 1 g/L) or high-glucose (HG, 5 g/L) conditions for 48 h. α-Tubulin served as the loading control. Quantitative protein levels of ITGB2 in BV2 and Müller cells ( n = 3 per group). B: Representative immunofluorescence images of retinal cryosections from control and streptozotocin (STZ)-induced diabetic mice, co-stained for HDGF (red) and the microglial marker IBA1 (green). Nuclei were counterstained with DAPI (blue). Assessment of immunofluorescence colocalization of HDGF and IBA1 in the two experimental groups (left, control; right, STZ). C: Single-cell RNA sequencing analysis of control and STZ-diabetic mouse retinas. UMAP plot showing the expression of Ncl (the gene encoding nucleolin) across all cells (upper). The microglia cluster is indicated. Dot plot illustrating the relative expression level and proportion of Ncl -expressing cells in microglia in the control and STZ groups (lower). D: Representative immunofluorescence images (left) and quantification of mean fluorescence intensity (MFI, right) for NCL (green) in BV2 cells cultured under LG or HG conditions for 48 h. Nuclei were counterstained with DAPI (blue). n = 3. Data are presented as mean ± standard deviation. Statistical significance was determined by the paired Student's t -test. * P < 0.05. Abbreviations: HDGF, hepatoma-derived growth factor; IBA1, ionized calcium-binding adaptor molecule 1; ns, not significant; STZ, streptozotocin.

Article Snippet: The immortalized murine microglial cell line (BV2) (Cat. #CRL-2469) was purchased from the American Type Culture Collection and cultured in DMEM/F12 medium (Gibco) supplemented with 10% FBS (Invitrogen), 1% glutamine (Invitrogen), and streptomycin (100 U/mL; Invitrogen) at 37 °C, 5% CO 2 .

Techniques: Expressing, Western Blot, Cell Culture, Control, Immunofluorescence, Staining, Marker, RNA Sequencing, Fluorescence, Standard Deviation, Derivative Assay, Binding Assay

Journal: Frontiers in Pharmacology

Article Title: Comprehensive profiling of Rhodiola rosea roots and corresponding products: phytochemical insights and modulation of neuroinflammation in BV2 microglial cell model

doi: 10.3389/fphar.2025.1608767

Figure Lengend Snippet: The influence of tested extracts (50 μg/mL) and rosavin (0.1–5 μM) on TNF-α secretion by LPS-stimulated microglia BV2. Data from three separate experiments assayed in duplicate are expressed as mean ± SEM. Dexamethasone (DEX, 20 μM) was used as a positive control. Absorbance values for all samples were expressed as percentages relative to the LPS-stimulated control (KST), which was set at 100%. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001 vs. stimulated control (KST), # statistically significant ( p < 0.001) vs. non-stimulated control (KNST).

Article Snippet: The immortalised murine microglial cell line BV2 (passages 1–4) was purchased from DSMZ–German Collection of Microorganisms and Cell Cultures GmbH.

Techniques: Positive Control, Control

Journal: Frontiers in Pharmacology

Article Title: Comprehensive profiling of Rhodiola rosea roots and corresponding products: phytochemical insights and modulation of neuroinflammation in BV2 microglial cell model

doi: 10.3389/fphar.2025.1608767

Figure Lengend Snippet: The influence of tested extracts (50 μg/mL) and rosavin (0.1–5 μM) on IL-6 secretion by LPS-stimulated microglia BV2. Data from three separate experiments assayed in duplicate are expressed as mean ± SEM. Dexamethasone (DEX, 20 μM) was used as a positive control. Absorbance values for all samples were expressed as percentages relative to the LPS-stimulated control (KST), which was set at 100%. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001 vs. stimulated control (KST), # statistically significant ( p < 0.001) vs. non-stimulated control (KNST).

Article Snippet: The immortalised murine microglial cell line BV2 (passages 1–4) was purchased from DSMZ–German Collection of Microorganisms and Cell Cultures GmbH.

Techniques: Positive Control, Control

Journal: Materials Today Bio

Article Title: Enhanced inhibition of neuronal ferroptosis and regulation of microglial polarization with multifunctional traditional Chinese medicine active ingredients-based selenium nanoparticles for treating spinal cord injury

doi: 10.1016/j.mtbio.2025.101758

Figure Lengend Snippet: TSIIA/TMP/APS@Se NPs regulate the polarization of BV2 cells and improve the inflammatory microenvironment to rescue PC12 cells in a co-culture system. (A–D) The ELISA quantitative analysis of the inflammatory cytokines in BV2 cells after different treatments. (E) Schematic illustration of the modulation of TSIIA/TMP/APS@Se NPs on BV2 polarization. (F) Protein expression images of iNOS and Arg-1 in BV2 cells after different treatment of nanoparticles. (G) Flow cytometry result of the BV2 cells to identify the M1 phenotype (F4/80, CD16/32 double positive) and M2 phenotype (F4/80, CD206 double positive) after incubation with F4/80, CD16/32, CD206. (H) Schematic illustration of a co-cultured system between PC12 cells and BV2 cells, indicating outcomes of diverse treatments with Se NPs. (I) The representative images of Live/Dead stains of PC12 cells after different treatments of the co-cultured system. Scale bar = 200 μm ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, and ns ( P > 0.05) suggested no statistical difference.

Article Snippet: The immortalized murine microglial cell line BV2 is commonly used as a surrogate for primary microglia in CNS research [ ].

Techniques: Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Incubation, Cell Culture

Journal: Cell Discovery

Article Title: Structural basis for allosteric agonism of human α7 nicotinic acetylcholine receptors

doi: 10.1038/s41421-025-00788-y

Figure Lengend Snippet: a Chemical structure of GAT is shown. Representative responses of α7 or α7 EM nAChR are shown for the application of ACh or GAT in bath solutions containing 2 mM Ca 2+ with a 10-s duration. b Effect of GAT on the release of inflammatory molecule IL-1β in LPS-stimulated microglial BV2 cells. The concentrations of LPS, GTS21, and MLA used in the experiment are 1 μg/mL, 100 μM, and 10 μM, respectively. Data are shown as means ± SEM ( n ≥ 3). P -values were calculated using an unpaired two-tailed Student’s t -test. * P < 0.05, ** P < 0.01. c Cryo-EM maps and ribbon representation of the complex in the open state. The Asn-linked carbohydrates and associated residues (Asn46, Asn90, and Asn133) are shown as sticks (green). d Cryo-EM maps and ribbon representation of the complex in a desensitized state. The Asn-linked carbohydrates and associated residues (Asn46, Asn90, and Asn133) are shown as sticks (green). GAT molecules are shown as spheres (yellow). e Plots of pore radius for α7 receptors along the pore axis. The α-carbon position of 0’ (Lys261) is set to zero. Channel pore radius was calculated using the HOLE program.

Article Snippet: The murine BV2 microglial cell line was purchased from Procell (CL-0493A).

Techniques: Two Tailed Test, Cryo-EM Sample Prep