Journal: The Journal of Biological Chemistry

Article Title: Cytosolic nucleic acid sensing triggers type I interferon activation via the Hippo kinase LATS1

doi: 10.1016/j.jbc.2026.111204

Figure Lengend Snippet: Pharmacological Inhibition of LATS impedes cytosolic nucleic acid sensing PRR activation of IFN-I. A , immunoblot analysis ( left panel ) and relative quantification ( right panel ) of IRF3 (S396) phosphorylation in MEF cells pretreated with vehicle control (DMSO) or LATS inhibitor (20 μM; 2.5 h) then transfected with pI:C or ISD (2 μg/ml each; 3 h). B , qPCR analysis of Ifnb mRNA in MEF cells pretreated with DMSO or LATS inhibitor as in A , followed by pI:C transfection (2 μg/ml; 3.5 h) ( left panel ) or ISD transfection (2 μg/ml; 4 h) ( right panel ). C and D , qPCR of Cxcl10, Ccl5, and Isg15 mRNA in MEF cells pretreated with DMSO or LATS1 inhibitor followed by pI:C transfection as in B , left panel ( C ) or ISD as in B , right panel ( D ). Statistical significance was determined using student’s t test (∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05).

Article Snippet: Forskolin, Vadimezan (DMXAA), and recombinant murine IFNβ were purchased from MedChemExpress.

Techniques: Inhibition, Activation Assay, Western Blot, Quantitative Proteomics, Phospho-proteomics, Control, Transfection

Journal: The Journal of Biological Chemistry

Article Title: Cytosolic nucleic acid sensing triggers type I interferon activation via the Hippo kinase LATS1

doi: 10.1016/j.jbc.2026.111204

Figure Lengend Snippet: LATS1 is required for IFN-I activation. A and B , qPCR analysis of Ifnb mRNA expression in WT and Lats1 −/− MEF cells infected with VSV (MOI 0.5) ( A ) or HSV-1 (MOI 0.1) for 24 h. C and D , qPCR analysis of Cxcl10 and Ccl5 mRNA in WT and Lats1 −/− MEF cells infected with VSV as in panel A ( C ) or HSV-1 as in panel B ( D ). E and F , immunoblot analysis ( left panels ) and relative quantification ( right panels ) of IRF3 (S396) phosphorylation in WT and Lats1 −/− MEF cells transfected with pI:C ( E ) or ISD ( F ) (2 μg/ml each) for the indicated times. G and H , qPCR analysis of Ifnb mRNA expression in WT and Lats1 −/− MEF cells transfected with pI:C ( G ) or ISD ( H ) (2 μg/ml each) for the indicated times. I and J , ELISA for IFN-β secretion in WT and Lats1 −/− MEF cells transfected with pI:C ( I ) or ISD ( J ) (2 μg/ml each; 6 h). K and L , qPCR analysis of Cxcl10, Ccl5, and Isg15 mRNA in WT and Lats1 −/− MEF cells transfected with pI:C ( K ) or ISD ( L ) (2 μg/ml each) for the indicated times. Statistical significance was determined using Student’s t test (∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05).

Article Snippet: Forskolin, Vadimezan (DMXAA), and recombinant murine IFNβ were purchased from MedChemExpress.

Techniques: Activation Assay, Expressing, Infection, Western Blot, Quantitative Proteomics, Phospho-proteomics, Transfection, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Biological Chemistry

Article Title: Cytosolic nucleic acid sensing triggers type I interferon activation via the Hippo kinase LATS1

doi: 10.1016/j.jbc.2026.111204

Figure Lengend Snippet: LATS1 operates at the level of TBK1. A , immunoblot analysis of LATS1 expression in HEK 293T cells treated with control siRNA or siRNAs targeting LATS1 for 24 h. B , IFN-β luciferase reporter assay in HEK 293T cells treated with control siRNA or siRNAs targeting LATS1 as in A , followed by transfection of plasmids encoding RIG-I, MAVS, STING, TBK1, and IRF3. C , co-immunoprecipitation and immunoblot of FLAG-LATS1 and HA-TBK1 co-transfected in HEK 293T cells. D and E , immunoblot analysis of TBK1-LATS1 interactions in MEF cells transfected with pI:C ( D ) or ISD ( E ) (2.5 μg/ml each; 2 h). Statistical significance was determined using student’s t test (∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05).

Article Snippet: Forskolin, Vadimezan (DMXAA), and recombinant murine IFNβ were purchased from MedChemExpress.

Techniques: Western Blot, Expressing, Control, Luciferase, Reporter Assay, Transfection, Immunoprecipitation

Journal: The Journal of Biological Chemistry

Article Title: Cytosolic nucleic acid sensing triggers type I interferon activation via the Hippo kinase LATS1

doi: 10.1016/j.jbc.2026.111204

Figure Lengend Snippet: LATS1 facilitates TBK1 activation and signaling events. A and B , immunoblot analysis ( left panels ) and relative quantification ( right panels ) of TBK1 (S172) phosphorylation in WT and Lats1 −/− MEF cells transfected with pI:C ( A ) or ISD ( B ) (2 μg/ml each) for the indicated times. C , immunoblot analysis of TBK1 (S172) phosphorylation in HEK 293T cells co-transfected with plasmids encoding WT TBK1 or kinase dead (KD) TBK1 (K38A) in the absence or presence of LATS1. D , IFN-β luciferase reporter assay in HEK 293T cells co-transfected with plasmids encoding TBK1 in the presence of WT LATS1 or kinase dead (KD) LATS1 (D846A). E , co-immunoprecipitation and immunoblot ( left panel ) and relative quantification ( right panel ) of WT LATS1-TBK1 and KD LATS1-TBK1 interactions in HEK 293T cells co-transfected with the indicated plasmids. F , co-immunoprecipitation and immunoblot ( left panel ) and relative quantification ( right panel ) of TBK1-IRF3 interactions in the absence or presence of LATS1 in HEK 293T cells co-transfected with the indicated plasmids. G and H , immunoblot analysis of TBK1-IRF3 interactions in WT and Lats1 −/− MEF cells transfected with pI:C ( G ) or ISD ( H ) (2 μg/ml each; 2.5 h). Statistical significance was determined using student’s t test (∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05).

Article Snippet: Forskolin, Vadimezan (DMXAA), and recombinant murine IFNβ were purchased from MedChemExpress.

Techniques: Activation Assay, Western Blot, Quantitative Proteomics, Phospho-proteomics, Transfection, Luciferase, Reporter Assay, Immunoprecipitation

Journal: The Journal of Biological Chemistry

Article Title: Cytosolic nucleic acid sensing triggers type I interferon activation via the Hippo kinase LATS1

doi: 10.1016/j.jbc.2026.111204

Figure Lengend Snippet: Model of Hippo kinase LATS1-dependent regulation of IFN-I in cytosolic nucleic acid sensing PRR signaling pathways. Under basal conditions, YAP operates as a steady-state negative regulator of TBK1-IRF3 signaling in part by associating with TBK1. Upon the detection of RNA or DNA virus genomes by distinct cytosolic PRRs, the Hippo kinase LATS1 undergoes phosphorylation to trigger the phospho-inhibition of YAP, abrogating its ability to antagonize TBK1. Activated LATS1 further associates with TBK1 and licenses TBK1 dependent signaling to IRF3, resulting in the subsequent induction of IFN-I. Figure created with BioRender.

Article Snippet: Forskolin, Vadimezan (DMXAA), and recombinant murine IFNβ were purchased from MedChemExpress.

Techniques: Protein-Protein interactions, Virus, Phospho-proteomics, Inhibition

Journal: Disease Models & Mechanisms

Article Title: The role of 25-hydroxycholesterol in the pathophysiology of brain vessel dysfunction associated with infection and cholesterol dysregulation

doi: 10.1242/dmm.052145

Figure Lengend Snippet: 25HC remodels cholesterol metabolism and function of brain endothelial cells. (1) CH25H upregulation was detected in SARS-CoV-2-associated ICH in zebrafish and human foetal brain tissue, as well as in hCMEC/D3 cells exposed to antiviral stimuli [poly(I:C) and IFNβ]. (2) 25HC treatment induced the downregulation of cholesterol synthesis enzymes at both mRNA and protein levels. In zebrafish and hCMEC/D3 cells, this downregulation had an additive effect when combined with pharmacological inhibition of HMGCR by ATV, leading to brain vessel rupture (ICH) in zebrafish and decreased barrier function and cell migration in hCMEC/D3 cells. (3) 25HC treatment in hCMEC/D3 cells also increased cholesterol efflux, which was associated with upregulation of ABCG1. (4) Internalisation of cholesterol into lipid droplets was not observed in 25HC-treated hCMEC/D3 cells. (5) The changes in cholesterol synthesis and efflux in hCMEC/D3 cells were associated with the depletion of plasma membrane-accessible cholesterol. (6) Cholesterol supplementation rescued the levels of accessible cholesterol and mitigated the decrease in barrier function and cell migration induced by 25HC in hCMEC/D3 cells.

Article Snippet: Cells were treated with high-molecular mass poly(I:C) (InvivoGen, tlrl-pic), human IFNβ (Tonbo Biosciences, 21-8699), murine IFNβ (R&D Systems, 12401-1), 25HC (Sigma-Aldrich, H1015), soluble cholesterol (Sigma-Aldrich, C4951), Staurosporine (Cell Guidance System, SM97), Mitomycin C (Roche, 10107409001) or ATV (Merck, SML3030), at concentrations and times stated in figure legends.

Techniques: Inhibition, Migration, Clinical Proteomics, Membrane