Journal: Nature communications

Article Title: Macrophage ILF3 promotes abdominal aortic aneurysm by inducing inflammatory imbalance in male mice.

doi: 10.1038/s41467-024-51030-4

Figure Lengend Snippet: Fig. 4 | Macrophage ILF3 contributes to Ang-II-induced inflammatory cell infiltration and the inflammatory response. A Representative Gene Ontology terms of differentially expressed genes (DEGs) enriched in ILF3+ macrophage clusters based on functional enrichment analysis. B Representative Kyoto Ency- clopedia of Genes and Genomes (KEGG) pathway data for macrophage DEGs enriched in ILF3+ vs ILF3-. C Immuno-staining of macrophage-derived MCP-1, TNF-α, IL-6, and IL-10 levels in AAA lesions of ILF3M-WT ApoE−/−and ILF3M-KO ApoE−/−mice treated with Ang II for four weeks (n = 6 per group; Scale bar=25 μm). D Plasma IL-6, TNF-α, MCP-1, IL10 levels in ILF3M-WT ApoE−/−and ILF3M-KO ApoE−/−mice treated with

Article Snippet: The resulting cell lysates and anti-ILF3 antibody (19887-1-AP, Proteintech) were mixed overnight in a rotary mixer at 4 °C.

Techniques: Functional Assay, Immunostaining, Derivative Assay, Clinical Proteomics

Journal: Nature communications

Article Title: Macrophage ILF3 promotes abdominal aortic aneurysm by inducing inflammatory imbalance in male mice.

doi: 10.1038/s41467-024-51030-4

Figure Lengend Snippet: Fig. 8 | Take home image/conceptual framework of the study. Under the AAA pathological environment, ILF3 was upregulated in aneurysmal macrophages. Macrophage ILF3 activation accelerates the degradation of p105 mRNA and increases nuclear p65 translocation, intensifying pro-inflammatory responses via activation of the NF-κB pathway. On the other hand, macrophage ILF3 may bind to Keap1 mRNA via the protein complex ILF3-eIF4A1 and amplifies Keap1 translation,

Article Snippet: The resulting cell lysates and anti-ILF3 antibody (19887-1-AP, Proteintech) were mixed overnight in a rotary mixer at 4 °C.

Techniques: Activation Assay, Translocation Assay

Journal: Scientific reports

Article Title: ILF2 and ILF3 are autoantigens in canine systemic autoimmune disease.

doi: 10.1038/s41598-018-23034-w

Figure Lengend Snippet: Figure 3. Screening the canine cohort for ILF3 and RBMX autoantibodies. A radio-ligand binding assay was used to screen for ILF3 (a) and RBMX (b) autoantibodies in dogs. The cutoff value for ILF3 was calculated from healthy controls as the mean + 5 SD. For RBMX, the mean + 7 SD was used as cutoff value because of a clear separation of negative and positive samples. All samples were analysed in duplicate. Autoantibody index = (sample value mean − negative control)/(positive control − negative control) * 100. NSDTR, Nova Scotia duck tolling retriever. IMRD ANAS, IMRD patients with speckled antinuclear antibodies (ANA) pattern. IMRD ANAH, IMRD patients with homogenous ANA pattern. IMRD ANAneg, IMRD patients without antinuclear antibodies. SRMA, steroid-responsive meningitis-arteritis.

Article Snippet: Human ILF3 cDNA in a pCMV6-Entry vector (RC214999, Origene) was used for in vitro transcription and translation of ILF3.

Techniques: Radio Ligand Binding Assay, Negative Control, Positive Control

Journal: Scientific reports

Article Title: ILF2 and ILF3 are autoantigens in canine systemic autoimmune disease.

doi: 10.1038/s41598-018-23034-w

Figure Lengend Snippet: Figure 5. Sera from ILF2- and ILF3-positive patients and commercial ILF2 and ILF3 antibodies display a speckled ANA pattern. Indirect immunofluorescence microscopy images of HEp-2 cells incubated with sera from an ILF2- and ILF3-positive IMRD patient (a), a commercial polyclonal ILF2 antibody (b); and a commercial polyclonal ILF3 antibody (c). (a) In the nucleoplasm, fine tiny speckles can be observed and mitotic cells have unstained chromatin mass (⇨). The nucleoli are not stained (→). (b,c) In the nucleoplasm, fine tiny speckles can be observed and mitotic cells have unstained chromatin mass (⇨). Nucleoli are stained in many of the cells (→) but not all.

Article Snippet: Human ILF3 cDNA in a pCMV6-Entry vector (RC214999, Origene) was used for in vitro transcription and translation of ILF3.

Techniques: Immunofluorescence, Microscopy, Incubation, Staining

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: Rat cerebrocortical cultures were treated with gp120 from HIV-1 strain BaL (200 pM) for 24 h in the presence or absence of different IFNβ concentrations (500 or 5,000 U/ml) or BSA/PBS (0.001% final concentration) as vehicle control. Following the incubation the cells were fixed and permeabilized, and neurons were immunolabeled for neuronal MAP-2 and NeuN while nuclear DNA was stained with H33342. Neuronal survival was assessed using fluorescence microscopy and cell counting as described in Methods. Values are mean ± s.e.m.; n = 2 independent experiments with 4–7 replicates and an average of 5,300 cells counted per condition; ***p < 0.001, **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Concentration Assay, Control, Incubation, Immunolabeling, Staining, Fluorescence, Microscopy, Cell Counting

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: Sagittal brains sections of HIVgp120tg and WT littermate mice previously treated with intranasal IFNβ or vehicle (veh) were immunolabeled for CCL4, neuronal MAP-2 or astrocytic GFAP. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was labeled with Hoechst (H) 33342. The fluorescence-labeled brain sections were analyzed using confocal laser-scanning microscopy. Representative images of cortex layer III are shown; scale bar, 50 μm.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Immunolabeling, Labeling, Fluorescence, Confocal Laser Scanning Microscopy

Journal: Scientific Reports

Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

doi: 10.1038/srep46514

Figure Lengend Snippet: ( a ) Cerebrocortical cultures from mice were prepared to either contain microglia, neurons and astrocytes (M + N + A) or were depleted of microglia (N + A) or neurons and microglia (A). Complete and depleted cell cultures were incubated with mIFNβ (5,000 U/ml) or BSA/PBS vehicle control for 0, 3, 6, 12 and 24 h and concentrations of CCL4 were measured in cell-free supernatants using a commercially available multiplex assay as described in Methods. Maximum concentrations were reached in samples of 12 to 24 h mIFNβ exposure and compared to vehicle-treated, baseline samples. Values are mean ± s.e.m.; n = 3 independent experiments; *p < 0.05, student’s t-test. ( b ) Microglia-depleted rat cerebrocortical cultures were exposed for 24 h to 50% cell-free conditioned media (CM) from human MDM in the presence or absence of human IFNβ (5,000 U/ml). MDM were previously stimulated for 24 h with HIV-1 gp120 BaL (MDM gp120 CM) or vehicle (MDM CM). Following the incubation the cells were fixed and permeabilized. Neurons were immunolabeled for neuronal MAP-2 and NeuN and nuclear DNA was stained with H33342. Neuronal survival was assessed using fluorescence microscopy and cell counting as described in Methods. Values are mean ± s.e.m.; n = 2 independent experiments, with 4–8 replicates and an average of 4,000 cells counted per condition; **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test.

Article Snippet: For visualization of CCL4, mouse brain sections were immunolabeled with primary antibodies for CCL4 (ProSci, Poway, CA, cat# 7227, 1:50) in combination with Ab against MAP-2 or GFAP for 24 h. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was stained with Hoechst (H) 33342.

Techniques: Incubation, Control, Multiplex Assay, Immunolabeling, Staining, Fluorescence, Microscopy, Cell Counting