Journal: bioRxiv

Article Title: Olfactory proteomics reveals the capacity of the HDAC1 inhibitor pyroxamide to halt the α-synuclein preformed fibrils-induced damage in nasal epithelial, microglial and dopaminergic neuronal cell lines

doi: 10.1101/2025.10.02.679944

Figure Lengend Snippet: In vitro experiments in dopaminergic MN9D cells representing the survival effect of α-syn PFFs after 6h of pyroxamide pre-treatment (A). Survival effect of α-syn PFFs after 3 and 6 hours of pyroxamide pre-treatment in olfactory epithelial T9243 cells (B). Survival effect of α-syn PFFs after 3 hours of pyroxamide pre-treatment in microglial BV2 cells upon α-syn PFFs-induced damage (C). Data were analyzed using one-way ANOVA. * indicates p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Similar conditions were used for the Dopaminergic Neuronal Cell Line (MN9D; Merck Millipore, SCC281), and the Mouse Microglial Cell Line (BV2; Cytion, 305156), seeded at 1 × 10^4 cells/well in 96-well plates.

Techniques: In Vitro

Journal: bioRxiv

Article Title: Olfactory proteomics reveals the capacity of the HDAC1 inhibitor pyroxamide to halt the α-synuclein preformed fibrils-induced damage in nasal epithelial, microglial and dopaminergic neuronal cell lines

doi: 10.1101/2025.10.02.679944

Figure Lengend Snippet: Survival analysis associated to pyroxamide pre-treatment 3 hours in dopaminergic neuronal (A) and microglial cells (B) upon oxidative stress conditions. Data were analyzed using one-way ANOVA. * indicates p < 0.05; ** p < 0.01.

Article Snippet: Similar conditions were used for the Dopaminergic Neuronal Cell Line (MN9D; Merck Millipore, SCC281), and the Mouse Microglial Cell Line (BV2; Cytion, 305156), seeded at 1 × 10^4 cells/well in 96-well plates.

Techniques:

Journal: Journal of Anesthesia and Translational Medicine

Article Title: MANF improves cognitive function and attenuates neuroinflammation in APP/PS1 transgenic mice through the TLR4/MYD88/NF-κB signaling pathway

doi: 10.1016/j.jatmed.2025.12.005

Figure Lengend Snippet: MANF inhibits Aβ1–42-induced neuroinflammation in BV2 cells via the TLR4/NF-κB signaling pathway. (A) Schematic diagram illustrates the experimental design for studying the effects of MANF intervention on Aβ1–42-induced neuroinflammation in BV2 cells. (B–D) ELISA analysis of the concentrations of inflammatory cytokines TNF-α, IL-6, and IL-1β in the supernatant of BV2 cell cultures (n = 4). MANF intervention significantly reduced the secretion of inflammatory cytokines induced by Aβ1–42. (E) Representative immunofluorescence images of the neuroinflammation marker IBA-1 (red) and nuclear staining with DAPI (blue) in BV2 cells. Scale bar = 100 μm. (F) Quantitative analysis of IBA-1-positive cells (n = 3) shows that MANF significantly decreased Aβ1–42-induced activation of BV2 cells. (G) Confocal images show increased nuclear translocation of p65 in Aβ-treated BV2 cells, which was attenuated by MANF treatment. p65 is shown in yellow and nuclei are stained with DAPI (blue) (n = 6). (H) Western blot analysis of TLR4/NF-κB pathway-related proteins, including TLR4, Myd88, PP65, and p-IKBα, in BV2 cells. (I–L) Quantitative analysis of the protein expression levels of TLR4, Myd88, PP65, and p-IKBα (n = 6). The data demonstrates that MANF significantly inhibited Aβ1–42-induced neuroinflammatory responses by downregulating the expression of TLR4/NF-κB pathway-related proteins. All data are presented as mean ± standard error of the mean (S.E.M.). Statistical significance is indicated as follows: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: To assess the regulatory influence of MANF on neuroinflammation and apoptosis, two distinct cell lines were utilized: HT22 mouse hippocampal neuronal cells and BV2 mouse microglial cells (Procell, China).

Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Marker, Staining, Activation Assay, Translocation Assay, Western Blot, Expressing

Journal: Journal of Anesthesia and Translational Medicine

Article Title: MANF improves cognitive function and attenuates neuroinflammation in APP/PS1 transgenic mice through the TLR4/MYD88/NF-κB signaling pathway

doi: 10.1016/j.jatmed.2025.12.005

Figure Lengend Snippet: MANF intervention inhibits apoptosis of HT22 cells in a co-culture system with Aβ1–42-induced BV2 cells. (A) Schematic diagram illustrates the experimental design of the co-culture system involving BV2 cells and HT22 cells. (B) rhMANF treatment improved the viability of HT22 cells (n = 4). (C) Representative immunofluorescence images of TUNEL staining in HT22 cells. TUNEL (green) and DAPI (blue) staining demonstrate that MANF significantly inhibited Aβ1–42-induced apoptosis in HT22 cells. Scale bar = 50 μm. (D) Quantitative analysis of TUNEL-positive HT22 cells (n = 4) further confirms the anti-apoptotic effects of MANF. (E–I) Western blot quantitative analysis of apoptosis-related proteins, including Bax, Bcl2, and Cleaved-caspase-3, in HT22 cells (n = 6). The results show that MANF upregulated the expression of the anti-apoptotic protein Bcl2 while downregulating the expression of pro-apoptotic proteins Bax and Cleaved-caspase-3. (J) Representative Western blot bands of Bax, Bcl2, and Cleaved-caspase-3 in HT22 cells support the regulatory effects of MANF on apoptosis-related protein expression. All data are presented as mean ± standard error of the mean (S.E.M.). Statistical significance is indicated as follows: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: To assess the regulatory influence of MANF on neuroinflammation and apoptosis, two distinct cell lines were utilized: HT22 mouse hippocampal neuronal cells and BV2 mouse microglial cells (Procell, China).

Techniques: Co-Culture Assay, Immunofluorescence, TUNEL Assay, Staining, Western Blot, Expressing

Journal: Journal of Inflammation Research

Article Title: Downregulation of Gbp2 Attenuates LPS-Induced Inflammation in BV2 Microglia Cells Through Inhibition of STAT1

doi: 10.2147/JIR.S531241

Figure Lengend Snippet: Downregulation of Gbp2 decreased inflammatory response and increased cell proliferation in LPS-stimulated BV2 cells. ( A ) BV2 cells were transfected with Gbp2 siRNA1 and Gbp2 siRNA2. RT-qPCR assay was performed to determine Gbp2 levels in BV2 cells. ( B–D ) BV2 cells were transfected with Gbp2 siRNA1 and Gbp2 siRNA2, followed by exposed to LPS. RT-qPCR and Western blot assays were conducted to determine Gbp2 levels in BV2 cells. ( E and F ) ELISA was employed to analyze IL-6 and TNF-α levels in the supernatant of BV2 cells. ( G ) Cell proliferation was determined by the EdU staining assay. **P<0.01 vs Blank group; ## P<0.01 vs LPS group.

Article Snippet: Mouse microglial BV2 cells (CL-0493) were sourced from Procell, and grown in DMEM medium containing 10% FBS and 1% penicillin-streptomycin (P/S).

Techniques: Transfection, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Staining

Journal: Journal of Inflammation Research

Article Title: Downregulation of Gbp2 Attenuates LPS-Induced Inflammation in BV2 Microglia Cells Through Inhibition of STAT1

doi: 10.2147/JIR.S531241

Figure Lengend Snippet: Downregulation of Gbp2 inhibited cell apoptosis in LPS-stimulated BV2 cells. BV2 cells were transfected with Gbp2 siRNA1 and Gbp2 siRNA2, followed by exposed to LPS. ( A ) IF staining assay was conducted to detect the expression of cleaved caspase 3 in BV2 cells. ( B ) Cell apoptosis was determined by the TUNEL staining assay. **P<0.01 vs Blank group; ## P<0.01 vs LPS group.

Article Snippet: Mouse microglial BV2 cells (CL-0493) were sourced from Procell, and grown in DMEM medium containing 10% FBS and 1% penicillin-streptomycin (P/S).

Techniques: Transfection, Staining, Expressing, TUNEL Assay

Journal: Journal of Inflammation Research

Article Title: Downregulation of Gbp2 Attenuates LPS-Induced Inflammation in BV2 Microglia Cells Through Inhibition of STAT1

doi: 10.2147/JIR.S531241

Figure Lengend Snippet: The anti-apoptotic and anti-inflammatory effects of Gbp2 deficiency is mediated by the TFs STAT1 and Irf1. ( A ) Network of Gbp2 and its transcription factors STAT1 and Irf1. ( B ) BV2 cells were transfected with STAT1 siRNA1 and STAT1 siRNA2. RT-qPCR assay was performed to determine STAT1 levels in BV2 cells. ( C ) BV2 cells were transfected with STAT1 siRNA1 and STAT1 siRNA2, followed by exposed to LPS. Cell apoptosis was determined by the TUNEL staining assay. ( D and E ) ELISA was employed to analyze IL-6 and TNF-α levels in the supernatant of BV2 cells. ( F and G ) Western blot assay was conducted to measure STAT1, Irf1 and Gbp2 levels in BV2 cells. **P<0.01 vs Blank group; ## P<0.01 vs LPS group.

Article Snippet: Mouse microglial BV2 cells (CL-0493) were sourced from Procell, and grown in DMEM medium containing 10% FBS and 1% penicillin-streptomycin (P/S).

Techniques: Transfection, Quantitative RT-PCR, TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay, Western Blot