bv2 mouse microglia cell lines  (AcceGen Biotechnology)

Journal: Frontiers in Immunology

Article Title: miR-511-3p dysregulation-mediated AKT3/USP8 signaling imbalance: a molecular bridge between neuroinflammation and PSCI

doi: 10.3389/fimmu.2026.1766326

Figure Lengend Snippet: Regulatory effects of miR-511-3p on the AKT3/USP8 in BV2 cells. (A–C) . Relative expression of miR-511-3p/AKT3/USP8 in BV2 cell model. (D) . miR-511-3p mimic promoted miR-511-3p expression. (E) . pcDNA3.1-AKT3 promoted AKT3 expression, but miR-511-3p mimic inhibited AKT3 expression, while this inhibition was rescued by pcDNA3.1-AKT3. (F) . pcDNA3.1-AKT3 inhibited USP8 expression, but miR-511-3p mimic promoted USP8 expression, while this promotion was rescued by pcDNA3.1-AKT3. ** P < 0.01, *** P < 0.001, compared to mimic NC; ## P < 0.01, compared to pcDNA3.1; & P < 0.05, &&& P < 0.001, compared to miR-511-3p mimic.

Article Snippet: The mouse microglia cell line BV2 cells (Procell, China, CL-0697) were resuscitated, seeded in six-well plates, added with DMEM containing 10% fetal bovine serum (FBS).

Techniques: Expressing, Inhibition

Journal: Frontiers in Immunology

Article Title: miR-511-3p dysregulation-mediated AKT3/USP8 signaling imbalance: a molecular bridge between neuroinflammation and PSCI

doi: 10.3389/fimmu.2026.1766326

Figure Lengend Snippet: miR-511-3p inhibited inflammation by targeting AKT3/USP8 in the cell model. (A) The expression level of IL-1β was regulatory by miR-511-3p/AKT3/USP8. (B) The expression level of IL-6 was regulatory by miR-511-3p/AKT3/USP8. (C) The expression level of TNF-α was regulatory by miR-511-3p/AKT3/USP8. *** P < 0.001, compared to BV2; ## P < 0.01, ### P < 0.001, compared to BV2+OGD/R; & P < 0.05, && P < 0.01, &&& P < 0.001, compared to BV2+OGD/R+miR-511-3p mimic.

Article Snippet: The mouse microglia cell line BV2 cells (Procell, China, CL-0697) were resuscitated, seeded in six-well plates, added with DMEM containing 10% fetal bovine serum (FBS).

Techniques: Expressing

Journal: Communications Biology

Article Title: Targeting microglial inflammation in Parkinson’s disease: irisin activates PAFAH1B1-RAGE ubiquitination and TFEB-dependent autophagy to alleviate neurodegeneration

doi: 10.1038/s42003-025-09389-7

Figure Lengend Snippet: A – D Effects of irisin on the inflammatory response of LPS-activated BV2 cells. Treatment with 100 ng/mL irisin for 12 h reduces the number of activated cells, alleviates cell swelling, and suppresses the inflammatory response of microglial cells. PBS vs LPS, * p < 0.05; LPS + Irisin vs. LPS, * p < 0.05. E and F Expression of LC3II/I and P62 in PD mice and BV2 cells. Both the PD group and the LPS-treated group exhibit decreased LC3II/I expression and increased P62 expression. E Control vs. PD, * p < 0.05; PD + Irisin vs. PD, * p < 0.05. F PBS vs. LPS, * p < 0.05; LPS + Irisin vs, LPS, * p < 0.05. G Restoration of autophagic flux in BV2 cells by irisin after labeling autophagosomes with RFP-GFP-LC3. Scale bars: 10 µm. PBS vs. LPS, * p < 0.05; LPS + Irisin vs LPS, * p < 0.05. H TEM results showing the number of autolysosomes in the LPS group and the restoration effect of irisin treatment. PBS vs LPS, * p < 0.05; LPS + Irisin vs. LPS, * p < 0.05.

Article Snippet: The mouse microglia cell line BV2 (CL-0493, Procell, Wuhan, China) was cultured and passaged in DMEM (KeGene Bio, China) medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin.

Techniques: Expressing, Control, Labeling

Journal: Communications Biology

Article Title: Targeting microglial inflammation in Parkinson’s disease: irisin activates PAFAH1B1-RAGE ubiquitination and TFEB-dependent autophagy to alleviate neurodegeneration

doi: 10.1038/s42003-025-09389-7

Figure Lengend Snippet: Expression of RAGE mRNA and protein in the substantia nigra of PD mice, PD + Irisin mice, and the control group. A Control vs. PD, not significant; PD + Irisin vs. PD, not significant. B Control vs. PD, * p < 0.05; PD + Irisin vs. PD, * p < 0.05. Expression of RAGE mRNA and protein in BV2 microglial cells in the LPS-stimulated group, PBS control group, and irisin-treated group. C PBS vs. LPS, not significant; LPS + Irisin vs. LPS, not significant. D PBS vs. LPS, * p < 0.05; LPS + Irisin vs. LPS, * p < 0.05. E Prediction of 35 candidate ubiquitin E3 ligases using the Ubibrowser database. F Venn diagram screening the intersection of ubiquitination E3 ligases with interacting and differentially expressed mRNAs. Expression of PAFAH1B1 mRNA and protein in the substantia nigra of PD mice ( G ) and the LPS-induced BV2 microglial model ( H ). G Control vs. PD, * p < 0.05; PD + Irisin vs. PD, * p < 0.05. H PBS vs. LPS, * p < 0.05; LPS + Irisin vs. LPS, * p < 0.05.

Article Snippet: The mouse microglia cell line BV2 (CL-0493, Procell, Wuhan, China) was cultured and passaged in DMEM (KeGene Bio, China) medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin.

Techniques: Expressing, Control, Ubiquitin Proteomics

Journal: Communications Biology

Article Title: Targeting microglial inflammation in Parkinson’s disease: irisin activates PAFAH1B1-RAGE ubiquitination and TFEB-dependent autophagy to alleviate neurodegeneration

doi: 10.1038/s42003-025-09389-7

Figure Lengend Snippet: A Effect of overexpression of PAFAH1B1 on RAGE protein expression and the reversal effect of MG132. NC vs. oe-PAFAH1B1, * p < 0.05; oe-PAFAH1B1 + MG132 vs. oe-PAFAH1B1, * p < 0.05. B Binding energy analysis of PAFAH1B1 and RAGE. C and D Co-IP and immunolocalization results confirming the binding interaction between PAFAH1B1 and RAGE. Scale bars: 10 µm. E Protein half-life analysis of RAGE when PAFAH1B1 is inhibited. F : Ubiquitination level of RAGE upon co-treatment with si-PAFAH1B1 and MG132. G Increase in the ubiquitination level of RAGE in BV2 cells after irisin addition. H Reversal of irisin-induced ubiquitination of RAGE by combining irisin with si-PAFAH1B1.

Article Snippet: The mouse microglia cell line BV2 (CL-0493, Procell, Wuhan, China) was cultured and passaged in DMEM (KeGene Bio, China) medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin.

Techniques: Over Expression, Expressing, Binding Assay, Co-Immunoprecipitation Assay, Ubiquitin Proteomics

Journal: Communications Biology

Article Title: Targeting microglial inflammation in Parkinson’s disease: irisin activates PAFAH1B1-RAGE ubiquitination and TFEB-dependent autophagy to alleviate neurodegeneration

doi: 10.1038/s42003-025-09389-7

Figure Lengend Snippet: A and B Prediction of ubiquitination modification sites of RAGE using the database GPS-uber, showing potential ubiquitination modification at K61 and K169 sites, which are highly conserved among the three species. C Effects of single-point mutation plasmids (RAGE-K61R, K169R) and the 2KR plasmid on the ubiquitination level of RAGE. K61R and K169R partially attenuate the ubiquitination level, while 2KR attenuates almost all of it. D Evidence that PAFAH1B1 regulates RAGE ubiquitination through the K48 chain. E – H Effects of PAFAH1B1 and RAGE on autophagy and inflammation in BV2 cells. si-PAFAH1B1 reverses the irisin-induced enhancement of autophagic flux and the inhibitory effect on pro-inflammatory markers, while overexpression of PAFAH1B1 mimics the effects of irisin treatment. LPS vs. LPS + Irisin, * p < 0.05; LPS + Irisin vs. LPS + Irisin + si-PAFA, * p < 0.05; LPS + Irisin + si-PAFA vs. LPS + oe-PAFA, * p < 0.05; LPS vs. LPS + oe-PAFA, * p < 0.05; LPS + Irisin vs. LPS + oe-PAFA + oe-RAGE, * p < 0.05.

Article Snippet: The mouse microglia cell line BV2 (CL-0493, Procell, Wuhan, China) was cultured and passaged in DMEM (KeGene Bio, China) medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin.

Techniques: Ubiquitin Proteomics, Modification, Mutagenesis, Plasmid Preparation, Over Expression

Journal: Communications Biology

Article Title: Targeting microglial inflammation in Parkinson’s disease: irisin activates PAFAH1B1-RAGE ubiquitination and TFEB-dependent autophagy to alleviate neurodegeneration

doi: 10.1038/s42003-025-09389-7

Figure Lengend Snippet: A Expression of lysosomal markers TFEB and LAMP1 in the substantia nigra of PD mice and LPS-induced BV2 microglial cells. Control vs. PD, * p < 0.05; PD + Irisin vs. PD, * p < 0.05. B The nuclear and cytoplasmic expression of TFEB in BV2 cells treated with LPS and LPS combined with irisin was detected by WB. PBS vs. LPS, * p < 0.05; LPS + Irisin vs. LPS, * p < 0.05. C Immunofluorescence was used to detect the expression level of NLRP3 in the brain tissue of PD. Scale bars: 20 µm. Western blot analysis of NLRP3 inflammasome components (NLRP3, ASC, cleaved Caspase-1) in PD mice ( D ) and LPS-treated BV2 cells ( E ). E Control vs. PD, * p < 0.05; PD + Irisin vs. PD, * p < 0.05. F PBS vs. LPS, * p < 0.05; LPS + Irisin vs. LPS, * p < 0.05. F The morphology and expression of microglial marker IBA-1 in the brain tissue of the PD model were examined by IHC staining. G Immunofluorescence was used to detect the subcellular localization of LAMP1 and NLRP3 after LPS and irisin treatment. Scale bars: 20 µm. H The effect of autophagy intervention on the expression level of NLRP3 after LPS and irisin treatment was detected by WB. LPS vs. LPS + Irisin + MG132, * p < 0.05; LPS + Irisin + MG132 vs. LPS + Irisin + 3-MA, * p < 0.05; LPS + Irisin + 3-MA vs. LPS + Irisin, * p < 0.05; LPS vs. LPS + Irisin, * p < 0.05. I Behavioral assays were conducted to detect the behavioral changes in the PD model after overexpression of irisin and NLRP3. PD vs. PD + Irisin, * p < 0.05; PD + Irisin vs. PD + Irisin + NLRP3, * p < 0.05. J ELISA was used to detect the expression levels of inflammatory factors in each treatment group. PD vs. PD + Irisin, * p < 0.05; PD + Irisin vs. PD + Irisin + NLRP3, * p < 0.05. K The molecular mechanism diagram of irisin regulating autophagolysosomal damage recovery through PAFAH1B1-mediated ubiquitination of RAGE and improving the progression of PD.

Article Snippet: The mouse microglia cell line BV2 (CL-0493, Procell, Wuhan, China) was cultured and passaged in DMEM (KeGene Bio, China) medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin.

Techniques: Expressing, Control, Immunofluorescence, Western Blot, Marker, Immunohistochemistry, Over Expression, Enzyme-linked Immunosorbent Assay, Ubiquitin Proteomics