Journal: Cell Communication and Signaling : CCS

Article Title: Trem2 regulates microglial migratory responses via type I interferon signaling during photoreceptor degeneration

doi: 10.1186/s12964-025-02261-5

Figure Lengend Snippet: Decreased IFN-I signaling was observed in Trem2 -deficient microglia. ( A , B ) Gene set enrichment analysis (GSEA) using gene sets from Gene Ontology Biological Process (GOBP) ( A ) and Kyoto Encyclopedia of Genes and Genomes (KEGG) ( B ) databases shows decreased expression of genes associated with “type I interferon (IFN-I) signaling” (at P18) and “phagosome” (at P21) in Trem2 −/− :rd10 retina. The criteria for GSEA are set at|NES| > 1, p < 0.05 and FDR q < 0.25. ( C ) A heatmap generated to visualize the expression of interferon-stimulated genes (ISGs) in rd10 and Trem2 −/− :rd10 retina. ( D - F ) Relative mRNA expression of selected ISGs, including Cxcl10 ( D ), Ifitm3 ( E ), Ifnar1 ( F ), in purified Trem2 −/− :rd10 and rd10 microglia. n = 3, unpaired Students’ t test. ( G ) Representative images of IBA1 and IFITM3 co-staining in Trem2 −/− :rd10 and rd10 retina at P15, P18, P21 and P30. The white arrows and white squares indicate double positive cells. ( H ) Quantification of IFITM3/IBA1 fluorescence intensity ratio in IBA1 + cells indicates decreased IFITM3 expression in Trem2 −/− :rd10 microglia. n = 3, unpaired Students’ t test. Data are curated and presented as described in Fig.

Article Snippet: IFN-α/β receptor I (IFNAR1) neutralizing antibody (#16-5945-85, ThermoFisher Scientific, CA, USA) [ ] and control IgG antibody (#16-4714-82, ThermoFisher) were administrated by intravitreal injection of 2 μL at the day when MNU was injected.

Techniques: Expressing, Generated, Purification, Staining, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Trem2 regulates microglial migratory responses via type I interferon signaling during photoreceptor degeneration

doi: 10.1186/s12964-025-02261-5

Figure Lengend Snippet: IFN-I signaling was activated in mouse retina with microglia-specific Trem2 overexpression. ( A ) Relative mRNA expression of Trem2 in Tmem119 CreERT2 : Rosa26 CAG − LSL−Trem2 microglia with and without tamoxifen administration. n = 3, unpaired Students’ t test. ( B , C ) Representative images and quantification of TREM2 protein expression in microglia of two groups. The white arrows indicate TREM2-expression cells. n = 3, unpaired Students’ t test. ( D , E ) Representative images and quantification of IFITM3 expression in Vehicle + Control, Vehicle + MNU-3d and Tamoxifen + MNU-3d group. The white arrows indicate double positive cells. n = 3, one-way ANOVA with Tukey’s test. ( F , G ) Representative images and quantification of Phospho-STAT1 (p-STAT1) expression in the above three groups. The white arrows indicate double positive cells. n = 3, one‐way ANOVA with Tukey’s test. ( H - J ) Relative mRNA expression of selected ISGs, including Cxcl10 ( H ), Ifitm3 ( I ), Ifnar1 ( J ), in purified microglia from Vehicle + Control, Vehicle + MNU-3d and Tamoxifen + MNU-3d mouse retinas. n = 3, one‐way ANOVA with Tukey’s test. Data are curated and presented as described in Fig.

Article Snippet: IFN-α/β receptor I (IFNAR1) neutralizing antibody (#16-5945-85, ThermoFisher Scientific, CA, USA) [ ] and control IgG antibody (#16-4714-82, ThermoFisher) were administrated by intravitreal injection of 2 μL at the day when MNU was injected.

Techniques: Over Expression, Expressing, Control, Purification

Journal: Science Advances

Article Title: Pregnancy enhances antiviral immunity independent of type I IFN but dependent on IL-17–producing γδ + T cells in the nasal mucosa

doi: 10.1126/sciadv.ado7087

Figure Lengend Snippet: Kinetics of ( A ) IFN-α and ( B ) IFN-β concentrations measured in lung homogenates by ELISA (50 PFU; n = 3 to 5). ( C ) Kinetics of pulmonary viral load in Ifnar1 −/− mice measured by MDCK plaque assay (50 PFU; n = 5 to 10). ( D ) AM frequency and ( E ) absolute numbers measured in lung homogenates by flow cytometry (50 PFU; n = 5 to 11). ( F ) Frequency of pulmonary apoptotic AMs (CD45.2 + SiglecF + CD11c + Casp3 + ) at various time points post-IAV infection (50 PFU; n = 4 to 5). ( G ) Extracellular acidification rate (ECAR) and ( H ) oxygen consumption rate (OCR) of AMs by Seahorse assay before and after 24 hours of infection with HIN1 (MOI of 1) ( n = 8 to 10 mice pooled and plated in replicates). O, oligomycin; F, fluorocarbonyl cyanide phenylhydrazone; R/A, antimycin A/rotenone. ( I ) Naïve AMs derived from the BAL fluid of nonpregnant and pregnant mice infected in vitro with IAV (MOI of 1) for 24 hours and viral Ns1 expression quantified by RT-qPCR ( n = 8 to 10 mice pooled and plated in triplicates). ( J ) Naïve AMs from the BAL fluid of nonpregnant and pregnant mice adoptively transferred intratracheally (i.t.) (5 × 10 4 cells per 50 μl) into Rag1 −/− recipient mice and infected intranasally (i.n.) 2 hours posttransfer (50 PFU IAV). Created with BioRender.com . ( K ) Pulmonary viral load of Rag1 −/− recipient mice 3 days postinfection by MDCK plaque assay ( n = 5). ( L ) Murine AM depletion via intranasal administration of 70 μl of control (CTL) or clodronate liposome 2 days before IAV infection (50 PFU). Created with BioRender.com . ( M ) Pulmonary viral load 1 day post-IAV infection measured by MDCK plaque assay following AM depletion ( n = 6 to 10). Data are presented as mean ± SEM. with * P < 0.05, ** P < 0.01, and **** P < 0.0001. Data were analyzed using two-way ANOVA followed by Sidak’s multiple comparison [(A) to (F) and (M)], one-way ANOVA followed by Holm- Šidák’s multiple comparisons test [(G) and (H)], or unpaired Student’s t test [(I) and (K)].

Article Snippet: Eight- to 10-week-old female WT, Rag1 −/− (B6.129S7- Rag1 tm1Mom/J ), IFN-α/β receptor deficient ( Ifnar1 −/− ) (B6.129S2- Ifnar1 tm1Agt /Mmjax), and T cell receptor delta chain deficient ( TCR δ −/− ) (B6.129P2- Tcrd tm1Mom /J) C57BL/6 mice were purchased from the Jackson Laboratory housed, and bred with male C57BL/6 or BALB/c mice at the animal facility of the Research Institute of the McGill University Health Centre.

Techniques: Enzyme-linked Immunosorbent Assay, Plaque Assay, Flow Cytometry, Infection, Derivative Assay, In Vitro, Expressing, Quantitative RT-PCR, Control, Comparison