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SULF A-dependent modulation of microglial activation and arginine metabolism. ( A ) RT-PCR analysis of cytokine and microglial marker gene expression following 24 h treatment. ( B ) CCL2 secretion quantified by ELISA at 48 h (n = 12). ( C ) Flow cytometric analysis of surface marker expression <t>(CD40,</t> CD86, MHC II, CD200R, TREM2) on untreated microglia (CTRL) or cells treated with 10 µg/mL SULF A for 24 h, expressed as mean fluorescence intensity (MFI; n = 6). ( D ) Schematic of iNOS/ARG metabolic pathways. ( E ) Quantification of nitric oxide (NO) release in culture supernatants by Griess assay at 48 h (n = 12). ( F ) Frequency of microglial cells expressing ARG, iNOS, or both, after 48 h of SULF A treatment (n = 6); ( G–H ) Representative FACS plots showing ARG⁺, iNOS⁺, and ARG⁺iNOS⁺ populations in untreated and SULF A-treated microglia. Data represent mean ± SEM. Statistical significance determined by paired t-test (P < 0.05; **P < 0.001; ***P < 0.0001)
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SULF A-dependent modulation of microglial activation and arginine metabolism. ( A ) RT-PCR analysis of cytokine and microglial marker gene expression following 24 h treatment. ( B ) CCL2 secretion quantified by ELISA at 48 h (n = 12). ( C ) Flow cytometric analysis of surface marker expression <t>(CD40,</t> CD86, MHC II, CD200R, TREM2) on untreated microglia (CTRL) or cells treated with 10 µg/mL SULF A for 24 h, expressed as mean fluorescence intensity (MFI; n = 6). ( D ) Schematic of iNOS/ARG metabolic pathways. ( E ) Quantification of nitric oxide (NO) release in culture supernatants by Griess assay at 48 h (n = 12). ( F ) Frequency of microglial cells expressing ARG, iNOS, or both, after 48 h of SULF A treatment (n = 6); ( G–H ) Representative FACS plots showing ARG⁺, iNOS⁺, and ARG⁺iNOS⁺ populations in untreated and SULF A-treated microglia. Data represent mean ± SEM. Statistical significance determined by paired t-test (P < 0.05; **P < 0.001; ***P < 0.0001)
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A. and C. Absolute number of B220 + B cells in the bone marrow (one femur and one tibia ( n = 5-8 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two-tailed t -test (mean ± SEM). ns, not significant B . and D . Absolute number of B220 + B cells in the spleen ( n = 5-8 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two-tailed t -test (mean ± SEM). ns, not significant E. and G. Splenic B cells cultured with indicated cytokines for 96 h and stained for IgG1, IgE, IgG2b, or IgG3. ( n = 5-9 mice per genotype, where each data point represents a single mouse). Isotype switching frequency was normalized to two wild-type mice for each experiment. Significance was determined by a one-way ANOVA with Tukey’s correction (mean ± SEM). F. and H. Representative flow cytometry plots showing splenic B cells cultured with <t>anti-CD40</t> and IL-4 for 96 h and stained for IgG1.
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A. and C. Absolute number of B220 + B cells in the bone marrow (one femur and one tibia ( n = 5-8 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two-tailed t -test (mean ± SEM). ns, not significant B . and D . Absolute number of B220 + B cells in the spleen ( n = 5-8 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two-tailed t -test (mean ± SEM). ns, not significant E. and G. Splenic B cells cultured with indicated cytokines for 96 h and stained for IgG1, IgE, IgG2b, or IgG3. ( n = 5-9 mice per genotype, where each data point represents a single mouse). Isotype switching frequency was normalized to two wild-type mice for each experiment. Significance was determined by a one-way ANOVA with Tukey’s correction (mean ± SEM). F. and H. Representative flow cytometry plots showing splenic B cells cultured with <t>anti-CD40</t> and IL-4 for 96 h and stained for IgG1.
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SULF A-dependent modulation of microglial activation and arginine metabolism. ( A ) RT-PCR analysis of cytokine and microglial marker gene expression following 24 h treatment. ( B ) CCL2 secretion quantified by ELISA at 48 h (n = 12). ( C ) Flow cytometric analysis of surface marker expression (CD40, CD86, MHC II, CD200R, TREM2) on untreated microglia (CTRL) or cells treated with 10 µg/mL SULF A for 24 h, expressed as mean fluorescence intensity (MFI; n = 6). ( D ) Schematic of iNOS/ARG metabolic pathways. ( E ) Quantification of nitric oxide (NO) release in culture supernatants by Griess assay at 48 h (n = 12). ( F ) Frequency of microglial cells expressing ARG, iNOS, or both, after 48 h of SULF A treatment (n = 6); ( G–H ) Representative FACS plots showing ARG⁺, iNOS⁺, and ARG⁺iNOS⁺ populations in untreated and SULF A-treated microglia. Data represent mean ± SEM. Statistical significance determined by paired t-test (P < 0.05; **P < 0.001; ***P < 0.0001)

Journal: Journal of Neuroinflammation

Article Title: Microglial clearance, neuroprotection and cognitive recovery via a novel synthetic sulfolipid in Alzheimer’s disease

doi: 10.1186/s12974-025-03634-w

Figure Lengend Snippet: SULF A-dependent modulation of microglial activation and arginine metabolism. ( A ) RT-PCR analysis of cytokine and microglial marker gene expression following 24 h treatment. ( B ) CCL2 secretion quantified by ELISA at 48 h (n = 12). ( C ) Flow cytometric analysis of surface marker expression (CD40, CD86, MHC II, CD200R, TREM2) on untreated microglia (CTRL) or cells treated with 10 µg/mL SULF A for 24 h, expressed as mean fluorescence intensity (MFI; n = 6). ( D ) Schematic of iNOS/ARG metabolic pathways. ( E ) Quantification of nitric oxide (NO) release in culture supernatants by Griess assay at 48 h (n = 12). ( F ) Frequency of microglial cells expressing ARG, iNOS, or both, after 48 h of SULF A treatment (n = 6); ( G–H ) Representative FACS plots showing ARG⁺, iNOS⁺, and ARG⁺iNOS⁺ populations in untreated and SULF A-treated microglia. Data represent mean ± SEM. Statistical significance determined by paired t-test (P < 0.05; **P < 0.001; ***P < 0.0001)

Article Snippet: Surface markers were assessed by staining with anti-mouse CD11b-VioBlue, CD40-PE, CD86-FITC, CD200R-APC (Miltenyi Biotec), and TREM2-APC (R&D Systems) according to standard protocols.

Techniques: Activation Assay, Reverse Transcription Polymerase Chain Reaction, Marker, Gene Expression, Enzyme-linked Immunosorbent Assay, Expressing, Fluorescence, Griess Assay

A. and C. Absolute number of B220 + B cells in the bone marrow (one femur and one tibia ( n = 5-8 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two-tailed t -test (mean ± SEM). ns, not significant B . and D . Absolute number of B220 + B cells in the spleen ( n = 5-8 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two-tailed t -test (mean ± SEM). ns, not significant E. and G. Splenic B cells cultured with indicated cytokines for 96 h and stained for IgG1, IgE, IgG2b, or IgG3. ( n = 5-9 mice per genotype, where each data point represents a single mouse). Isotype switching frequency was normalized to two wild-type mice for each experiment. Significance was determined by a one-way ANOVA with Tukey’s correction (mean ± SEM). F. and H. Representative flow cytometry plots showing splenic B cells cultured with anti-CD40 and IL-4 for 96 h and stained for IgG1.

Journal: bioRxiv

Article Title: The ERCC6L2-MRI-KU complex coordinates NHEJ at staggered DNA double-strand breaks

doi: 10.1101/2025.11.28.691009

Figure Lengend Snippet: A. and C. Absolute number of B220 + B cells in the bone marrow (one femur and one tibia ( n = 5-8 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two-tailed t -test (mean ± SEM). ns, not significant B . and D . Absolute number of B220 + B cells in the spleen ( n = 5-8 mice per genotype, where each data point is a single mouse). Significance was determined by an unpaired two-tailed t -test (mean ± SEM). ns, not significant E. and G. Splenic B cells cultured with indicated cytokines for 96 h and stained for IgG1, IgE, IgG2b, or IgG3. ( n = 5-9 mice per genotype, where each data point represents a single mouse). Isotype switching frequency was normalized to two wild-type mice for each experiment. Significance was determined by a one-way ANOVA with Tukey’s correction (mean ± SEM). F. and H. Representative flow cytometry plots showing splenic B cells cultured with anti-CD40 and IL-4 for 96 h and stained for IgG1.

Article Snippet: B cells were stimulated with LPS (Sigma, 5ug/mL), mouse recombinant IL-4 (10ng/mL, PeproTech), and agonist anti-CD40 antibody (0.5ug/mL, Miltenyi Biotec).

Techniques: Two Tailed Test, Cell Culture, Staining, Flow Cytometry