Journal: Journal of Lipid Research

Article Title: EBV promotes alveolar trabecula resorption via extracellular vesicle remodeling by group IIA secreted phospholipase A 2

doi: 10.1016/j.jlr.2026.101014

Figure Lengend Snippet: sPLA 2 -IIA hydrolyzes EV phospholipids and promotes bone resorption through LPAR1 and EP4 receptor. A: Electrospray ionization mass spectrometry of lipids from Akata LCL EVs after treatment with sPLA 2 -IIA (n = 3). Whole lipids were extracted, and the levels of both lysophospholipids and polyunsaturated fatty acids were analyzed. Relative values are shown. Values are presented as the mean ± SEM. Statistical significance was determined using student’s t test: ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. B: THP-1 cells were cultured with Akata LCL EVs treated with or without sPLA 2 -IIA for 24 h. Prior to administration, THP-1 cells were pretreated with or without 10 μM AM966 or 1 μM EP4 receptor antagonist 1. C: Representative images of H&E staining, immunostaining of CTSK, and TRAP staining of the mouse maxillary palatal region. PBS, sPLA 2 -IIA, Akata LCL EVs, and sPLA 2 -IIA-treated Akata LCL EVs were injected into the mandibular incisor region. EVs are treated with sPLA 2 -IIA and varespladib prior to injection. Capitals indicate T: tooth, P: periodontal ligament, and B: bone region. A representative example is shown from the analysis of three mice. D: Quantification of CTSK and TRAP staining in the tissue shown in C (n = 3 mice per group). Values in B and D are presented as the mean ± SD. Statistical significance was determined using student’s t test: ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: Akata LCL-derived EVs were added at a concentration corresponding to 5 μg of total protein per well and incubated at 37°C in a humidified atmosphere containing 5% CO 2 for 24 or 48 h. For inhibitor treatment, cells were pretreated with 10 μM AM966 (HY-15277, MedChemExpress) or 1 μM EP4 receptor antagonist 1 (HY-133123, MedChemExpress) 30 min prior to EV exposure.

Techniques: Mass Spectrometry, Cell Culture, Staining, Immunostaining, Injection

Journal: bioRxiv

Article Title: The COX2-PGE2-PKA Axis Suppresses Antiviral Immunity by Inhibiting mtDNA-Dependent STING Activation

doi: 10.64898/2026.04.03.716411

Figure Lengend Snippet: (A) Heatmap showing relative expression of COX2 and type I ISGs in mice treated with either vehicle (Veh) or doxorubicin (Doxo) (GEO: GSE223698). (B) Heatmap showing relative expression of COX2 and type I ISGs in proliferating cells (Prof) and senescence cells (Sen) (GEO: GSE196610). (C) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1) for either 24 or 48 hours. Cell lysates were subjected to RT-qPCR to assess mRNA levels of COX2 (n = 3). (D) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1) for either 24 or 48 hours in the presence or absence of 1 µM celecoxib (COX2i). Cell lysates were collected and analyzed by ELISA to measure extracellular PGE 2 levels (n = 3). (E) A schematic illustrates PGE 2 -cAMP-PKA signaling. (F) THP-1 macrophages were treated with 1 µM PGE 2 in the presence or absence of 1 µM EP4 inhibitor (EP4i) for 16 hours. Cell lysates were collected and analyzed by ELISA to measure intracellular cAMP levels (n = 3). (G) THP-1 macrophages were treated with 1 µM PGE 2 at indicated concentrations in the presence or absence of 1 µM EP4 inhibitor (EP4i) for 16 hours. Cell lysates were collected and subjected to immunoblotting with the indicated antibodies. (H) THP-1 macrophages expressing ISRE-Luciferase were mock-infected or infected with HSV-1 (MOI = 1) in the presence of either 1 µM PGE 2 or 1 µM forskolin and 50 µM IBMX. At 16 h.p.i, cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (I-J) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1), followed by 1 µM PGE 2 stimulation in the presence of either 1 µM EP4 inhibitor (EP4i) or 1 µM PKA inhibitor (PKAi). At 24 h.p.i, cell lysates were subjected to RT-qPCR to assess mRNA levels of IFNβ (n = 3) (I) , and cell supernatants were subjected to ELISA to measure secreted levels of IFNβ (n = 3) (J) . (K) THP-1 macrophages expressing ISRE-Luciferase were mock-infected or infected with HSV-1 (MOI = 1), followed by 1 µM PGE 2 stimulation in the presence of either 1 µM EP4 inhibitor (EP4i) or 1 µM PKA inhibitor (PKAi). At 16 h.p.i, cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (L) THP-1 macrophages were infected with HSV-1 (MOI = 1), followed by 1 µM PGE 2 stimulation in the presence of either 1 µM EP4 inhibitor (EP4i) or 1 µM PKA inhibitor (PKAi). At 16 h.p.i, cell lysates were collected and subjected to RT-qPCR to assess HSV-1 UL30 genomic abundance (n = 3). Data are presented as mean ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Sidak’s multiple comparisons test. P -values are indicated.

Article Snippet: For inhibition studies, cells were pre-incubated with EP4 antagonist ONO AE3-208 (Tocris 3565), PKA inhibitor BLU0588 (TargetMol, T60169), during the starvation period.

Techniques: Expressing, Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot, Luciferase, Reporter Assay, Activity Assay

Journal: bioRxiv

Article Title: The COX2-PGE2-PKA Axis Suppresses Antiviral Immunity by Inhibiting mtDNA-Dependent STING Activation

doi: 10.64898/2026.04.03.716411

Figure Lengend Snippet: (A) Proteomic workflow in HEK293 cells with doxycycline-inducible expression of PKA Cα wild-type (WT) or W197R mutant (MUT). (B) The top 8 significantly enriched KEGG pathways within the mitochondrial quality control category identified from the proteomic dataset. (C) Schematic of the mt-mKeima mitophagy reporter. (D) Representative Airyscan live-cell imaging of THP-1 macrophages expressing mt-mKeima sensor treated with 1 µM PGE 2 in the presence of either 1 µM EP4 inhibitor (EP4i) or 1 µM PKA inhibitor (PKAi) for 16 hours. Scale bar, 10 µm. (E) Quantification of mitolysosome numbers shown in (D) (n = 10). Data were quantified from one representative experiment of three. (F) Representative images of live-cell 4D lattice light sheet imaging on THP-1 macrophages treated as in (D) . Scale bar, 20 µm. (G-H) Quantification of net mitolysosome displacement (n = 10) (G) and average mitolysosome speed (n = 12) (H) from imaging in (F) . (I) THP-1 macrophages treated with PGE 2 at indicated concentrations in the presence or absence of 1 µM PKA inhibitor (PKAi) for 16 hours. Mitochondrial fractions were isolated and subjected to immunoblotting with indicated antibodies. Band intensities were quantified and normalized to COX4 expression for PINK1 (J) and pUB Ser65 (K) (n = 3). (L) Working model illustrating that PGE 2 induces mitophagy and mitochondrial biogenesis to enhance mitochondrial homeostasis in a EP4- and PKA-dependent manner. Data are presented as mean ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Sidak’s multiple comparisons test. p -values are indicated.

Article Snippet: For inhibition studies, cells were pre-incubated with EP4 antagonist ONO AE3-208 (Tocris 3565), PKA inhibitor BLU0588 (TargetMol, T60169), during the starvation period.

Techniques: Expressing, Mutagenesis, Control, Live Cell Imaging, Imaging, Isolation, Western Blot