murine microglial cell line bv2  (Procell Inc)

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 Cellular Neuroscience

Article Title: Structure matters: commensal Phocaeicola vulgatus lipopolysaccharide induces attenuated microglial activation and preserves neuronal integrity

doi: 10.3389/fncel.2026.1796397

Figure Lengend Snippet: Differential effects of E. coli and P. vulgatus LPS on BV2 microglial cells. (A) BV2 cell viability after stimulation with increasing concentrations of E. coli or P. vulgatus LPS (0.1–100 ng/mL) using the MTT assay. Unstimulated cells (NS) served as negative control. (B) Immunofluorescence analysis of microglial marker Iba-1 (red) to evaluate cell morphology and number following exposure to 100 ng/mL of LPS (scale bar = 100 μm). (C) The relative number of Iba 1 + cells from the experiments in (B) , normalized to NS (set at 100%). (D) Iba-1 immunofluorescence intensity was calculated as the mean area of the positive signal per cell. (E) Nitrite quantification and (F) western blot analysis of iNOS expression. Data are presented as mean ± SEM. Statistical significance was assessed by ordinary one-way ANOVA followed by Tukey’s multiple-comparison test. # p < 0.05; #### p < 0.0001 vs. NS; * p < 0.05; *** p < 0.001, **** p < 0.0001 vs. LPS.

Article Snippet: The murine BV2 microglial cell line (bladder carcinoma cell line-derived, viral oncogene-transformed; #305156) and the human microglial clone-3 cell line (HMC3; #300102) were purchased from Cytion Biosciences (Heidelberg, Germany).

Techniques: MTT Assay, Negative Control, Immunofluorescence, Marker, Western Blot, Expressing, Comparison

Journal: Frontiers in Cellular Neuroscience

Article Title: Structure matters: commensal Phocaeicola vulgatus lipopolysaccharide induces attenuated microglial activation and preserves neuronal integrity

doi: 10.3389/fncel.2026.1796397

Figure Lengend Snippet: Differential modulation of cytokine release and intracellular signaling in BV2 microglia. BV2 cells were treated with increasing concentrations (0.1, 1, 10, and 100 ng/mL) of LPS derived from E. coli or P. vulgatus . Unstimulated cells (NS) served as negative controls. (A) Cytokine levels (IL-1β, IL-6, and TNF-α) were quantified in the culture supernatants by using ELLA assay. (B) Activation of intracellular signaling pathways (STAT3, Nf-kb, ERK1/2) was assessed by western blotting. Data are presented as mean ± SEM. Statistical significance was assessed by ordinary one-way ANOVA followed by Tukey’s multiple-comparison test. # p < 0.05; ## p < 0.01; ### p < 0.001, #### p < 0.0001 vs. NS; * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001 vs. LPS.

Article Snippet: The murine BV2 microglial cell line (bladder carcinoma cell line-derived, viral oncogene-transformed; #305156) and the human microglial clone-3 cell line (HMC3; #300102) were purchased from Cytion Biosciences (Heidelberg, Germany).

Techniques: Derivative Assay, Activation Assay, Protein-Protein interactions, Western Blot, Comparison

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Propranolol reduced the expression of NLRP3 and IL-1β in BV2 cells subjected to OGD/R (A) Schematic of cell experiment steps. (B) Western blot analysis of NLRP3 and IL-1β in BV2 cells at 4, 6, 8, 12, and 24 h after OGD/R. (C and D) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 versus the control group. (E) Western blot analysis of NLRP3 and IL-1β in BV2 cells of different treatment groups. (F and G) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 and ## p < 0.01 versus the OGD/R group.

Article Snippet: The murine microglial cell line BV2 was obtained from Procell Life Science & Technology Co., Ltd. (Procell, Cat# CL-0493).

Techniques: Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Blockade of β2-AR reduced the expression of NLRP3 in BV2 cells subjected to OGD/R (A) Western blot analysis of the effects of dobutamine and betaxolol on NLRP3 and IL-1β expression in BV2 cells. (B and C) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the control group. (D) Western blot analysis of salmeterol and ICI118,551 on NLRP3 and IL-1β expression in BV2 cells. (E and F) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 and ## p < 0.01 versus the OGD/R group.

Article Snippet: The murine microglial cell line BV2 was obtained from Procell Life Science & Technology Co., Ltd. (Procell, Cat# CL-0493).

Techniques: Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Activation of β2-AR induced the expression of NLRP3 and IL-1β via the ERK1/2 MAPK signaling pathway (A) Western blot analysis of the effects of H-89 and U0126 on NLRP3 and IL-1β expression in BV2 cells. (B and C) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 versus the OGD/R group. (D) Western blot analysis of p -ERK1/2 expression in BV2 cells. (E) Quantitative analysis of p -ERK1/2 expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 versus the OGD/R group.

Article Snippet: The murine microglial cell line BV2 was obtained from Procell Life Science & Technology Co., Ltd. (Procell, Cat# CL-0493).

Techniques: Activation Assay, Expressing, Western Blot, Control

Journal: The Journal of Headache and Pain

Article Title: hsa_circ_0006168 drives microglial activation in TNC via miR-99b-5p/KDM6B axis to promote central sensitization in migraine

doi: 10.1186/s10194-026-02288-0

Figure Lengend Snippet: The effect of mmu_circ_0012878 on microglia in the TNC of mice and LPS-treated BV2 cells. A Representative immunofluorescence images of Iba1 (green) and CD206 (red) co-localization in the TNC following mmu_circ_0012878 knockdown ( n = 3). Scale bar = 20 μm. B Representative microglial morphology from each group in the areas marked by white boxes in A. Scale bar = 5 μm. C Qulification of Iba1+/CD206+ double-positive cells ( n = 3). D Representative immunofluorescence images showing co-localization of Iba1 (green) and CD86 (red) in the TNC following mmu_circ_0012878 knockdown ( n = 3). Scale bar = 20 μm. E Qulification of Iba1+/CD86+ double-positive cells ( n = 3). F Western blot analysis of KDM6B at different time points following 1 μg/ml LPS stimulation ( n = 3). G-H the relative expression of mmu_circ_0012878 and miR-99b-5p in LPS-stimulated BV2 cells were measured using qRT-PCR ( n = 4). Data are shown as the means ± SD. The p values were calculated using one-way ANOVA ( C-F ). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The murine BV2 microglial cell line was obtained from Procell (Wuhan, China).

Techniques: Immunofluorescence, Knockdown, Western Blot, Expressing, Quantitative RT-PCR

Journal: The Journal of Headache and Pain

Article Title: hsa_circ_0006168 drives microglial activation in TNC via miR-99b-5p/KDM6B axis to promote central sensitization in migraine

doi: 10.1186/s10194-026-02288-0

Figure Lengend Snippet: mmu_circ_0012878 acted as a miR-99b-5p sponge to modulate KDM6B expression. A qRT-PCR analysis confirmed the knockdown efficiency of mmu_circ_0012878 ( n = 3). B Relative expression levels of miR-99b-5p in each group were assessed by qRT-PCR ( n = 3). C Western blot analysis of KDM6B expression in BV2 cells ( n = 3). D Western blot analysis of PI3K/AKT pathway protein expression in BV2 cells following mmu_circ_0012878 knockdown ( n = 3). E Immunofluorescence staining showing KDM6B expression and morphological changes in microglia ( n = 3). Scale bar = 50 μm. F Western blot was performed to investigate the regulatory relationships among mmu_circ_0012878, miR-99b-5p, and KDM6B ( n = 3). G Expression of PI3K/AKT pathway proteins in LPS-treated BV2 cells co-transfected with LV-mmu_circ_0012878-RNAi and KDM6B overexpression plasmids ( n = 3). H-I Predicted binding sites among mmu_circ_0012878, miR-99b-5p, and KDM6B are shown schematically. The predicted interactions were validated using dual luciferase assays ( n = 3). Data are shown as the means ± SD. The p values were calculated using one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The murine BV2 microglial cell line was obtained from Procell (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Western Blot, Immunofluorescence, Staining, Transfection, Over Expression, Binding Assay, Luciferase

Journal: The Journal of Headache and Pain

Article Title: hsa_circ_0006168 drives microglial activation in TNC via miR-99b-5p/KDM6B axis to promote central sensitization in migraine

doi: 10.1186/s10194-026-02288-0

Figure Lengend Snippet: Silencing mmu_circ_0012878 attenuated LPS-induced M1 polarization while enhancing M2 polarization in BV2 cells. A The protein expression levels of iNOS and Arg-1 were examined by western blot in each group ( n = 3). B-C Flow cytometry was performed to analysis the MFl of CD86 and CD206 ( n = 3). D-E The quantification of cytokines from multiplex flow cytometry assay ( n = 3). F-H ELISA analysis of IL-6, IL-1β and IL-12p70 expression levels ( n = 4). Data are shown as the means ± SD. The p values were calculated using one-way ANOVA. * p < 0.05, *** p < 0.001

Article Snippet: The murine BV2 microglial cell line was obtained from Procell (Wuhan, China).

Techniques: Expressing, Western Blot, Flow Cytometry, Multiplex Assay, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Targeting Microglial CD49a Inhibits Neuroinflammation and Demonstrates Therapeutic Potential for Parkinson's Disease

doi: 10.1002/advs.202515138

Figure Lengend Snippet: In vitro Itga1 knockdown attenuates microglial hyperactivation. (A,B) Representative flow cytometry (FC) images with quantitative analysis demonstrating the efficiency of Itga1 knockdown in BV2 cells using si‐ITGA1‐1 # and si‐ITGA1‐2 # ( n = 3). (C,D) Representative blots with quantitative analysis demonstrating the efficiency of Itga1 knockdown in primary microglia using si‐ITGA1‐1 # and si‐ITGA1‐2 # ( n = 3). (E–G) Representative FC images with quantitative analysis of CD86 and CD206 expression in primary microglia ( n = 3). (H,J,K) Representative immunofluorescence images with quantitative analysis of CD49a and IBA1 expression in BV2 cells ( n = 3). CD49a, green fluorescence; IBA1, red fluorescence; DAPI, blue fluorescence. Scale bar: 50 µm for original and 20 µm for magnified images. (I,L) Representative immunofluorescence images with quantitative analysis of latex beads' phagocytosis in BV2 cells ( n = 3). Latex beads, green fluorescence; Phalloidin, red fluorescence; DAPI, blue fluorescence. White arrows point to latex beads phagocytosed by BV2 cells. (M,N) mRNA expression levels of Itga1 , proinflammatory genes ( Il1b , Il6 , and Tnf ), and anti‐inflammatory genes ( Il10 , Il13 , and Arg1 ) in primary microglia ( n = 3). Data are presented as means ± SEM with t ‐test, one‐way ANOVA, and two‐way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: The murine microglial BV2 cell line (CL‐0493; RRID: CVCL_0182) was obtained from Procell (Wuhan, China).

Techniques: In Vitro, Knockdown, Flow Cytometry, Expressing, Immunofluorescence, Fluorescence

Journal: Advanced Science

Article Title: Targeting Microglial CD49a Inhibits Neuroinflammation and Demonstrates Therapeutic Potential for Parkinson's Disease

doi: 10.1002/advs.202515138

Figure Lengend Snippet: In vitro microglial Itga1 knockdown alleviates mitochondrial damage and NLRP3 inflammasome activation via PGAM5. (A,B) Representative blots and quantification of NLRP3, COX2, iNOS, ASC, and PGAM5 protein expression levels in primary microglia ( n = 3). (C,D) Representative flow cytometry (FC) images and quantification of intracellular and mitochondrial reactive oxygen species (ROS) generation in primary microglia ( n = 3). (E,G) Representative fluorescence images and quantification of mitochondrial Δψm in BV2 cells ( n = 3). J‐monomers, green fluorescence; J‐aggregates, red fluorescence. Scale bar: 50 µm for original and 20 µm for magnified images. (F,H) Ultrastructural images and quantification of mitochondria in BV2 cells ( n = 5). Scale bar: 500 nm for original and 200 nm for magnified images. Black arrows represent mitochondria in BV2 cells. The green, yellow, and red in the statistical chart represent normal, damaged, and degenerated mitochondria, respectively. Data are presented as means ± SEM with one‐way and two‐way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: The murine microglial BV2 cell line (CL‐0493; RRID: CVCL_0182) was obtained from Procell (Wuhan, China).

Techniques: In Vitro, Knockdown, Activation Assay, Expressing, Flow Cytometry, Fluorescence

Journal: Advanced Science

Article Title: Targeting Microglial CD49a Inhibits Neuroinflammation and Demonstrates Therapeutic Potential for Parkinson's Disease

doi: 10.1002/advs.202515138

Figure Lengend Snippet: Microglial PGAM5 overexpression reverses anti‐inflammatory effects and mitochondrial homeostasis mediated by Itga1 knockdown. (A) Experimental workflow depicting the experimental procedure involving the establishment of stable OE‐PGAM5 BV2 cells, transfection with si‐ITGA1‐2 # , LPS/Nig stimulation protocol, and detection. (B,C) Representative blots and quantification of PGAM5 protein expression levels in OE‐PGAM5 or Vec‐PGAM5 BV2 cells ( n = 3). (D) The mRNA expression levels of PGAM5 in OE‐PGAM5 or Vec‐PGAM5 BV2 cells ( n = 3). (E,H–K) Representative blots and quantification of NLRP3, COX2, iNOS, ASC, and PGAM5 protein expression levels in OE‐PGAM5 or Vec‐PGAM5 BV2 cells ( n = 3). (F,G) Representative FC images and quantification of both cellular and mitochondrial ROS levels in OE‐PGAM5 or Vec‐PGAM5 BV2 cells ( n = 3). (L,N) Representative fluorescence images and quantification of mitochondrial Δψm in OE‐PGAM5 or Vec‐PGAM5 BV2 cells ( n = 3). J‐monomers, green fluorescence; J‐aggregates, red fluorescence. Scale bar: 10 µm. (M,O) Ultrastructural images and quantification of mitochondria in OE‐PGAM5 or Vec‐PGAM5 BV2 cells ( n = 5). Red arrows represent mitochondria. Scale bar: 200 nm. The green, yellow, and red in the statistical chart represent respectively normal, damaged, and degenerated mitochondria. (P) The mRNA transcript abundance of proinflammatory genes ( Il1b , Il6 , and Il18 ) in OE‐PGAM5 or Vec‐PGAM5 BV2 cells ( n = 3). Data are represented as means ± SEM with t ‐test, one‐way ANOVA, and two‐way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: The murine microglial BV2 cell line (CL‐0493; RRID: CVCL_0182) was obtained from Procell (Wuhan, China).

Techniques: Over Expression, Knockdown, Transfection, Expressing, Fluorescence