Journal: Scientific Reports

Article Title: Interferon-β and FTY720 ameliorate progressive CNS inflammation via SOCS1-associated astrocyte signaling

doi: 10.1038/s41598-026-45303-9

Figure Lengend Snippet: Glial cells respond to FTY720 and IFN-β treatment with cell type–specific modulation of inflammatory and regenerative programs. Primary cultures of murine glial cells were treated with vehicle, FTY720, or the combination of FTY720 and IFN-β overnight and then stimulated with IL-1β and TNF-ɑ for 4 h. qPCR of the indicated genes was performed in ( A-D ) microglia and ( n = 3 biological replicates) and ( E–F ) astrocytes (n = 6 biological replicates). ( G ) Astrocyte-conditioned medium (ACM) was generated as described in Fig. H. In brief, primary astrocytes were activated with IL-1β (10 ng/mL) and TNF-α (5 ng/mL) for 24 h, followed by treatment with FTY720/IFN-β and either the control peptide or pJAK2(1001–1013) (20 µM) for 6 h in FCS-free medium. The medium was replaced the next day and collected after an additional 24 h. The harvested ACM was added to activated astrocytes under the respective conditions for subsequent qPCR analysis. qPCR was performed of indicated genes (n = 6 biological replicates). ( H ) Migration assay was performed as described in Fig. H, and the number of migrated monocytes was measured after 3 h ( n = 5 biological replicates). ( I ) Activated human microglia (HMC3 cells; n = 6) and ( J ) human astrocytes (Human astrocytes Catalog #1800, ScienCell; n = 3) were treated with vehicle, FTY720 and vehicle, or the combination of FTY720 and IFN-β overnight and then stimulated with IL-1β and TNF-ɑ for 4 h. qPCR of the indicated genes was performed. All qPCR results are displayed by fold change in mRNA expression. P-values of p < 0.05 were considered significant and determined by one-way ANOVA followed by Tukey´s multiple-comparisons test.

Article Snippet: For further in vitro experiments we used the human microglia cell line HMC3 (American Type Culture Collection [ATCC], Manassas, VA, USA; CRL-3304) and primary human astrocytes (ScienCell Research Laboratories, Carlsbad, CA, USA; Human Astrocytes, Catalog #1800).

Techniques: Generated, Control, Migration, Expressing

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: Nature Communications

Article Title: A common 19 bp APOE enhancer deletion is protective against Alzheimer’s disease in African Americans

doi: 10.1038/s41467-026-68808-3

Figure Lengend Snippet: a Schematic of the luciferase experimental design and constructs. The APOE 3′UTR ( APOE 3′UTR only) plus ~400 bp of downstream DNA containing the 19 bp deletion region and SPI1 binding site were cloned into a psiCheck2.2 dual luciferase reporter construct. b Renilla:Firefly luciferase expression data in HMC3 cells after delivery of APOE SPI1 WT sequence ( p = 0.0016 relative to the APOE 3′UTR only) or delivery of the 19 bp deletion [ p = 0.8252; one-way ANOVA, n = 12 samples/group (3 biological replicates, 4 technical replicates), ± SD]. c –e HMC3 gene expression following SPI1 overexpression. c Effect of SPI1 overexpression on c APOE expression ( p = 0.3831, F = 1.789, unpaired two-tailed t-test, n = 12 control and 12 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM), d APOC1 expression ( p = <0.0001, F = 2.987, unpaired two-tailed t-test, n = 11 control and 12 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM), and e lncRNA ENSG00000280087 expression ( p = 0.0170, F = 5.414, unpaired two-tailed t-test, n = 12 control and 10 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM). f – h HMC3 gene expression following Aβ 1:42 treatment. f Effect of Aβ 1:42 treatment on APOE expression ( p = 0.2098, F = 1.243, unpaired two-tailed t-test, n = 12 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM), g APOC1 expression ( p = 0.0015, F = 4.657, unpaired two-tailed t-test, n = 11 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM) and h, lncRNA ENSG00000280087 expression ( p = 0.0255, F = 2.502, unpaired two-tailed t-test, n = 12 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM) in HMC3 cells.

Article Snippet: HMC3 human microglia cell lines (ATCC CRL-3304) were grown in EMEM media and seeded at 3.34 × 10 4 cells plated in 12-well plates for luciferase assays.

Techniques: Luciferase, Construct, Binding Assay, Clone Assay, Expressing, Sequencing, Gene Expression, Over Expression, Two Tailed Test, Control, Derivative Assay