Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Interleukin-1β-induced arthritis involves chondrocyte oxiapoptophagy

doi: 10.4196/kjpp.25.279

Figure Lengend Snippet: (A, B) IL-1β increases ROS production in chondrocytes. Scale bar = 100 μm. (C–E) The expression of inflammatory mediators, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nitric oxide (NO), and prostaglandin E 2 (PGE 2 ), are upregulated in chondrocytes stimulated with IL-1β. Values are presented as mean ± SD. DCF-DA, dichlorfluorescein-diacetate.

Article Snippet: The production of PGE 2 in the chondrocytes stimulated with IL-1β was measured using a PGE 2 Parameter Assay Kit (R&D System), according to the manufacturer’s instruction.

Techniques: Expressing

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) Schematic of the antiviral property of cGAS/STING-mediated type I IFN during HSV-1 infection. (B) Representative images of bone marrow-derived macrophages infected with GFP-expressing HSV-1 (MOI = 1) for 24 hours in the presence or absence of 1 µM PGE 2 . (C) Bone marrow-derived macrophages were infected with HSV-1 (MOI = 1) in the presence or absence of 1 µM PGE 2 . At 24 h.p.i, cell lysates were collected and subjected to qPCR to quantify HSV-1 UL30 genomic abundance (n = 3). Data are shown as ΔΔCt values. (D) Schematic of the COX2/PGE 2 signaling axis. (E) Bone marrow-derived macrophages were infected with HSV-1 (MOI = 1) in the presence or absence of 1 µM celecoxib. At 24 h.p.i, cell lysates were collected and subjected to qPCR to quantify HSV-1 UL30 genomic abundance (n = 3). Data are shown as ΔΔCt values. (F) Schematic illustrates experimental design of bulk RNA-seq workflow in THP-1 macrophages under four conditions: vehicle (Veh), PGE 2 alone (PGE 2 ), infected with HSV-1 for 16 hours in the absence (HSV-1+Veh) or presence of exogenous PGE 2 (HSV-1+PGE 2 ) to study mechanisms in which PGE 2 controls innate immunity in response to HSV-1 infection. (G-H) Gene set enrichment analysis (GSEA) of RNA-seq data comparing HSV-1 + PGE2 versus HSV-1 + Veh. (G) Tables summarize enriched gene sets and associated statistics. (H) Representative enrichment plots for the indicated pathways. See also Fig. S1E. NES, normalized enrichment score; FDR, false discovery rate. (I) THP-1 macrophages expressing ISRE-Luciferase were mock-infected and infected with HSV-1 (MOI = 1) in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (J) THP-1 macrophages were mock-infected and infected with HSV-1 (MOI = 1) in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, cell lysates were collected and subjected to RT-qPCR to measure mRNA levels of representative type I ISGs (n = 3). Data are shown ΔΔCt values. (K) THP-1 macrophages were mock-infected and infected with HSV-1 (MOI = 1) in the presence or absence of 1 µM PGE 2 . At 24 h.p.i, supernatants were collected and analyzed by ELISA to measure secreted levels of IFNβ (n = 3). All experiments were performed with three independent biological replicates and repeated at least twice with reproducible results. Data are presented as mean ± s.e.m. Statistical significance was determined by unpaired, two-tailed Student’s t-test (C, E) or one-way ANOVA followed by Sidak’s multiple comparisons test (I-K); p- values are indicated.

Article Snippet: Concentration of extracellular PGE 2 was measured using the Prostaglandin E 2 ELISA Kit – monoclonal (Cayman, 514010) according to the manufacturer’s instructions.

Techniques: Infection, Derivative Assay, Expressing, RNA Sequencing, Luciferase, Reporter Assay, Activity Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Two Tailed Test

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) A schematic illustrates the activation of STING, TBK1, and IRF3 in response to double-stranded DNA (dsDNA) in the cytosol by cGAS. (B-D) THP-1 macrophages were mock-infected or infected with HSV-1 in the presence or absence of 1 µM PGE 2 . At the indicated h.p.i, cell lysates were collected and subjected to immunoblotting with the indicated antibodies (B) . Band intensity of phosphorylated TBK1 (C) and IRF3 (D) was quantified and normalized to total TBK1 and IRF3, respectively (n = 3). (E) THP-1 macrophages expressing ISRE-Luciferase were transfected with viral DNA at indicated concentrations in the presence or absence of 1 µM PGE 2 for 16 hours. Cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (F) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1) for the indicated times. Cell lysates were collected and subjected to immunoblotting with the indicated antibodies. Samples transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) were included as controls. (G) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1). At 16 h.p.i, cell lysates were collected and subjected to qPCR to assess levels of total mt-D-loop and mt-ND1 regions (n = 3). (H) Representative Airyscan live-cell imaging of THP-1 macrophages mock-infected or infected with HSV-1 for 3 hours. MitoTracker DeepRed (magenta): mitochondria, PicoGreen (yellow): DNA. Scale bars, 2 µm. (I) Quantification of cytosolic PicoGreen in THP-1 macrophages mock-infected or infected with HSV-1 (MOI = 1) for 3 hours (n = 10). (J-K) THP-1 macrophages were infected with HSV-1 in the presence or absence of the mtDNA replication inhibitor, ddC. At 24 h.p.i, cell supernatants were collected and subjected to ELISA to measure levels of secreted IFNβ (n = 3) (J) , and cell lysates were collected and subjected to qPCR to quantify HSV-1 UL30 genomic abundance (n = 3) (K) . (L) THP-1 macrophages expressing ISRE-Luciferase were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) in the presence or absence of 1 µM PGE 2. At 16 hours post stimulation, cell lysates were collected and subjected to a luciferase reporter assay to assess ISRE promoter activity (n = 3). (M) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siTFAM) in the presence or absence of 1 µM PGE 2 . At 16 hours post-stimulation, cell lysates were collected and subjected to RT-qPCR to assess the mRNA levels of representative type I ISGs (n = 3). All experiments were performed with three independent biological replicates and repeated at least twice with reproducible results. Data are presented as mean ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Sidak’s multiple comparisons test ( C-E, L-M ) or unpaired, two-tailed Student’s t-test ( G, I-K ). p -values are indicated.

Article Snippet: Concentration of extracellular PGE 2 was measured using the Prostaglandin E 2 ELISA Kit – monoclonal (Cayman, 514010) according to the manufacturer’s instructions.

Techniques: Activation Assay, Infection, Western Blot, Expressing, Luciferase, Transfection, Reporter Assay, Activity Assay, Live Cell Imaging, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Two Tailed Test

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: Concentration of extracellular PGE 2 was measured using the Prostaglandin E 2 ELISA Kit – monoclonal (Cayman, 514010) according to the manufacturer’s instructions.

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: Concentration of extracellular PGE 2 was measured using the Prostaglandin E 2 ELISA Kit – monoclonal (Cayman, 514010) according to the manufacturer’s instructions.

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

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) THP-1 macrophages were treated with 1 µM PGE 2 alone, 1 µM forskolin and 50 µM IBMX alone, or infected with HSV-1 (MOI = 1) in the presence or absence of PGE 2 or forskolin/IBMX. At 16 h.p.i, cell lysates were collected and subjected to immunoblotting with indicated antibodies. (B) Representative Airyscan live-cell imaging of THP-1 macrophages transfected with either control siRNA (siCTRL) or TFAM siRNA (siRNA) in the absence or presence of 1 µM PGE 2 for 16 hours. Scale bars, 2 µm. (C) Quantification of cytosolic PicoGreen from images in (B) (n = 10). Data were quantified from one representative experiment of three. (D) Schematic showing the workflow to quantify cytosolic mitochondrial DNA using qPCR. (E) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or TFAM siRNA (siRNA) in the presence or absence of 1 µM PGE 2 for 16 hours. Cytosol fractions were isolated and subjected to qPCR to assess the presence of mt-Dloop and mt-ND1 regions (n = 3). (F) THP-1 macrophages were mock-infected or infected with HSV-1 (MOI = 1) in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, cytosol fractions were isolated and subjected to qPCR to assess the presence of mt-Dloop and mt-ND1 regions (n = 3). (G) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or PINK1 siRNA (siPINK1), followed by mock or HSV-1 infection in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, cytosol fractions were isolated and subjected to qPCR to assess the presence of mt-Dloop and mt-ND1 regions (n=3). (H) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or PINK1 siRNA (siPINK1), followed by mock or HSV-1 infection in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, whole cell lysates were collected and subjected to RT-qPCR to assess mRNA levels of IFNβ (n = 3). (I) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or PINK1 siRNA (siPINK1), followed by mock or HSV-1 infection in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, whole cell lysates were collected and subjected to qPCR to assess HSV-1 UL30 genomic abundance (n=3). All experiments were performed with three independent biological replicates and repeated at least twice with reproducible results. 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: Concentration of extracellular PGE 2 was measured using the Prostaglandin E 2 ELISA Kit – monoclonal (Cayman, 514010) according to the manufacturer’s instructions.

Techniques: Infection, Western Blot, Live Cell Imaging, Transfection, Control, Isolation, Quantitative RT-PCR

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) Network of mitochondrial quality control proteins interacting with wild-type (WT) and mutant (MUT) PKA Cα with BFDR σ; 0.2. Interactors are colored by log 2 fold change (WT/MUT spectral counts) with WT-specific interactors in dark purple and MUT-specific interactors in dark green. Edges to the central bait (yellow) represent interactions detected in this study. (B) Whole cell lysates of THP-1 macrophages were subjected to pulldown using 8-AHA-cAMP (RIα), Rp-8-AHA-cAMPS (holoenzyme), or HaloLink resin (control), followed by immunoblotting with indicated antibodies. (C-D) THP-1 macrophages were stimulated with 1 µM PGE 2 for 6 hours, followed by incubation with 1 µM cycloheximide for the indicated times. Cell lysates were collected and subjected to immunoblotting with the indicated antibodies. Representative blots from three independent experiments are shown (C) . Band intensities for STOML2 were quantified and normalized to HSP90 intensity (n = 3) (D) . (E-G) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or STOML2 siRNA (siSTOML2), followed by treatment with PGE 2 at indicated concentrations for 16 hours. Mitochondrial fractions were isolated and subjected to immunoblotting with the indicated antibodies (E) . Band intensities for PINK1 (F) and pUB Ser65 (G) were quantified and normalized to COX4 intensity (n = 3). (H) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or STOML2 siRNA (siSTOML2), followed by mock or HSV-1 infection in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, cytosol fractions were isolated and subjected to qPCR to assess the presence of mt-Dloop and mt-ND1 regions (n=3). (I) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or STOML2 siRNA (siSTOML2), followed by mock or HSV-1 infection in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, whole cell lysates were collected and subjected to RT-qPCR to assess mRNA levels of IFNβ (n=3). (J) THP-1 macrophages were transfected with either scramble siRNA (siCTRL) or STOML2 siRNA (siSTOML2), followed by mock or HSV-1 infection in the presence or absence of 1 µM PGE 2 . At 16 h.p.i, whole cell lysates were collected and subjected to qPCR to assess HSV-1 UL30 genomic abundance (n=3). All experiments were performed with three independent biological replicates and repeated at least twice with reproducible results. 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: Concentration of extracellular PGE 2 was measured using the Prostaglandin E 2 ELISA Kit – monoclonal (Cayman, 514010) according to the manufacturer’s instructions.

Techniques: Control, Mutagenesis, Western Blot, Incubation, Transfection, Isolation, Infection, Quantitative RT-PCR

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) HEK293 cells expressing STOML2-3xFlag were treated with 1 µM forskolin and 50 µM IBMX for the indicated times. Whole cell lysates were subjected to immunoprecipitation using DYKDDDDK Fab-Trap agarose and immunoblotting with the indicated antibodies. (B) Top: domain architecture of STOML2 and predicted PKA phosphorylation sites (Scansite 4.0; PhosphoSitePlus). Bottom: ClustalW alignment showing evolutionary conservation surrounding STOML2 Ser29. (C) Representative structural model of the interaction between the active pocket of PKA Cα and STOML2 N-terminal (aa 18 to 37) (ipTM = 0.9). Gray: PKA Cα, green: PKA Cα residue D167, yellow: STOML2, magenta: STOML2 residue S29. (D) HEK293 cells expressing either STOML2 wild-type (WT) or S29A mutant (MUT) were treated with 1 µM forskolin and 50 µM IBMX for 6 hours. Cell lysates were subjected to immunoprecipitation using DYKDDDDK Fab-Trap agarose and immunoblotting with the indicated antibodies. (E) Schematic illustrating the generation of THP-1 macrophages expressing either STOML2 wild-type (WT) or S29A mutant (MUT) via lentivirus transduction. (F) THP-1 macrophages expressing either STOML2 wild-type (WT) or S29A mutant (MUT) were stimulated with 1 µM PGE 2 for 16 hours. Cell lysates were subjected to immunoprecipitation using DYKDDDDK Fab-Trap agarose and immunoblotting with the indicated antibodies. (G) Representative Airyscan live-cell imaging of THP-1 macrophages expressing either STOML2 wild-type (WT) or S29A mutant (MUT) together with the mt-mKeima sensor were stimulated with 1 µM PGE 2 for 16 hours. Scale bar, 10 µm. (H) Quantification of mitolysosome numbers from (G) (n = 10). (I) THP-1 macrophages expressing either STOML2 wild-type (WT) or S29A mutant (MUT) were stimulated with 1 µM PGE 2 for 16 hours. Mitochondrial fractions were collected and subjected to immunoblotting with indicated antibodies. (J) A schematic illustrating that PKA activation promotes STOML2 phosphorylation at Ser29 which subsequently induces PINK1-mediated mitophagy in response to PGE 2 stimulation. All experiments were performed with three independent biological replicates and repeated at least twice with reproducible results. 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: Concentration of extracellular PGE 2 was measured using the Prostaglandin E 2 ELISA Kit – monoclonal (Cayman, 514010) according to the manufacturer’s instructions.

Techniques: Expressing, Immunoprecipitation, Western Blot, Phospho-proteomics, Residue, Mutagenesis, Transduction, Live Cell Imaging, Activation Assay

Journal: bioRxiv

Article Title: Aerobic exercise prevents the loss of endogenous pain modulation in male and female rats with traumatic brain injury

doi: 10.64898/2026.03.31.714901

Figure Lengend Snippet: The effect of the α 2 -adrenoceptor (α 2 -AR) antagonist, atipamezole (ATZ) on the intact DCN response of naïve male and female Sprague Dawley rats. The DCN response was blocked in both male and female naïve rats after treatment with ATZ when compared to VEH-treated naïve rats. A three-way mixed repeated measures ANOVA revealed significant main effects of drug condition (F(1,20)=18.34, p=3.63 x 10 -4 , □2=0.48), and time post-capsaicin (F(7,140)=33.31, p=6.51 x 10 -27 , □2=0.63). There were also significant interactions of drug condition by time post-capsaicin (F(7,140)=18.32, p=3.66 x 10 -17 , □2=0.48), and sex by drug condition by time post-capsaicin (F(7,140)=4.13, p=3.74 x 10 -4 , □2=0.17). Open arrow - PGE-2; i.pl. [1 μg/50 μl]. Closed arrow - Capsaicin; i.pl. [10 μg/10 μl]. Data are presented as mean + standard error of the mean. (n = 6 rats per cohort).

Article Snippet: Prostaglandin E2 (PGE-2) [1μg/50μl], intraplantar ( i.pl. ) (14010, Cayman Chemicals, MI, USA) a principal mediator of inflammation and pain hypersensitivity.

Techniques:

Journal: bioRxiv

Article Title: Aerobic exercise prevents the loss of endogenous pain modulation in male and female rats with traumatic brain injury

doi: 10.64898/2026.03.31.714901

Figure Lengend Snippet: Resolution of the acute phase of pain after TBI exposed the chronic phase of pain involving the failure of descending control of nociception (DCN) from 49 DPI ( and ) and up to 180 DPI ( and ) in both male and female TBI+Sed rats. In contrast the DCN response was intact in both male and female TBI+Ex rats at both 49 and 180 DPI . A four-way mixed repeated measures ANOVA revealed significant main effects of exercise (F(1,20)=2974.00, p=3.13 x 10 -23 , □2=0.99), time post-capsaicin (F(6,120)=120.52, p=2.20 x 10 -48 , □2=0.86) and time point (49 or 180 DPI) (F(1,20)=5.16, p=0.0343, □2=0.21). There was also a significant interaction of time post-capsaicin by exercise (F(6,120)=119.26, p=3.77 x 10 -48 , □2=0.86). ⋇ = TBI+Ex CT hindpaw vs. TBI+Sed CT hindpaw (p < 0.0001) by Bonferroni’s posthocs. Open arrow - PGE-2; i.pl. [1 μg/50 μl]. Closed arrow - Capsaicin; i.pl. [10 μg/10 μl]. Data are presented as mean + standard error of the mean. (n = 6 rats per cohort).

Article Snippet: Prostaglandin E2 (PGE-2) [1μg/50μl], intraplantar ( i.pl. ) (14010, Cayman Chemicals, MI, USA) a principal mediator of inflammation and pain hypersensitivity.

Techniques: Control

Journal: bioRxiv

Article Title: Aerobic exercise prevents the loss of endogenous pain modulation in male and female rats with traumatic brain injury

doi: 10.64898/2026.03.31.714901

Figure Lengend Snippet: The effect of the α 1 -adrenoceptor (α 1 -AR) antagonist, prazosin (PRZ) on the intact DCN response in female TBI+Ex rats 49 and 180 DPI . On day 49 of testing, half female TBI+Ex rats were randomly selected and given a systemic injection of PRZ or saline, the vehicle (VEH) ( and ) after the test stimulus (PGE-2). The DCN response was blocked in all female TBI+Ex rats treated with PRZ on day 49 post-TBI . For assessment at 180 DPI, a cross over design was adopted. Female TBI+Ex rats that had been treated with PRZ on day 49 post-TBI were treated with the VEH at 180 DPI and the TBI+Ex rats treated with the VEH on day 49 after injury were treated with PRZ at 180 DPI. The DCN response was also blocked in all female TBI+Ex rats treated with PRZ on day 180 post-TBI . VEH had no effect on the DCN response in female TBI+Ex rat at either timepoint ( and ). At 49 DPI a four-way mixed repeated measures ANOVA revealed significant main effects of drug condition (F(1,10)=1076.66, p=1.63 x 10 -11 , □2=0.99), and time post-capsaicin (F(7,70)=108.30, p=5.55 x 10 -35 , □2=0.92). There was also a significant interaction of time post-capsaicin by drug condition (F(7,70)=105.89, p=1.13 x 10 -34 , □2=0.91). At 180 DPI a four-way mixed repeated measures ANOVA revealed significant main effects of drug condition (F(1,10)=1459.41, p=3.60 x 10 -12 , □2=0.99), and time post-capsaicin (F(7,70)=137.94, p=2.31 x 10 -38 , □2=0.93). There was also a significant interaction of time post-capsaicin by drug condition (F(7,70)=134.92, p=4.74 x 10 -38 , □2=0.93). ⋇ = TBI+Ex+PRZ versus TBI+Ex+VEH (p < 0.0001) by Bonferroni’s posthocs. Open arrow - PGE-2; i.pl. [1 μg/50 μl]. Closed arrow - Capsaicin; i.pl. [10 μg/10 μl]. Data are presented as mean + standard error of the mean. (n = 6 rats per cohort).

Article Snippet: Prostaglandin E2 (PGE-2) [1μg/50μl], intraplantar ( i.pl. ) (14010, Cayman Chemicals, MI, USA) a principal mediator of inflammation and pain hypersensitivity.

Techniques: Injection, Saline

Journal: bioRxiv

Article Title: Aerobic exercise prevents the loss of endogenous pain modulation in male and female rats with traumatic brain injury

doi: 10.64898/2026.03.31.714901

Figure Lengend Snippet: The effect of the 5-HT 7 receptor antagonist, SB-267790 (SB-26), on the intact DCN response in male TBI+Ex rats 49 DPI . On the day of testing, half of the male TBI+Ex rats were randomly selected and given either an intrathecal injection of SB-26 (10 μg/10 μl, i.t.) or saline, the vehicle (VEH) after the test stimulus (PGE-2). The DCN response remained intact in male TBI+Ex rats treated with SB-26 at 49 DPI . VEH had no effect on the DCN response in male TBI+Ex rats. A four-way mixed repeated measures ANOVA revealed significant main effects of time post-capsaicin (F(7,70)=84.92, p=1.17 x 10 -31 , □2=0.90). Post hoc analysis reveals no significant difference in pain thresholds between TBI+Ex+SB-26 and TBI+Ex+VEH male rats. Data are presented as mean + standard error of the mean. (n = 6 rats per cohort).

Article Snippet: Prostaglandin E2 (PGE-2) [1μg/50μl], intraplantar ( i.pl. ) (14010, Cayman Chemicals, MI, USA) a principal mediator of inflammation and pain hypersensitivity.

Techniques: Injection, Saline

Journal: bioRxiv

Article Title: Aerobic exercise prevents the loss of endogenous pain modulation in male and female rats with traumatic brain injury

doi: 10.64898/2026.03.31.714901

Figure Lengend Snippet: The effect of the α 1 -adrenoceptor (α 1 -AR) antagonist, prazosin (PRZ) on the intact DCN response in male TBI+Ex rats 49 and 180 DPI . On day 49 of testing, half male TBI+Ex rats were randomly selected and given a systemic injection of PRZ or saline, the vehicle (VEH) ( and ) after the test stimulus (PGE-2). The DCN response was blocked in all male TBI+Ex rats treated with PRZ on day 49 post-TBI . For assessment at 180 DPI, a cross over design was adopted. Male TBI+Ex rats that had been treated with PRZ on day 49 post-TBI were treated with the VEH at 180 DPI and the TBI+Ex rats treated with the VEH on day 49 after injury were treated with PRZ at 180 DPI. The DCN response was also blocked in all male TBI+Ex rats treated with PRZ on day 180 post-TBI . VEH had no effect on the DCN response in male TBI+Ex rat at either timepoint ( and ). A four-way mixed repeated measures ANOVA comparing both male and female TBI+Ex PRZ and VEH treated rats at 49 DPI revealed significant main effects of drug condition (F(1,20)=1789.08, p=4.83 x 10 -21 , □2=0.99), and time post-capsaicin (F(7,140)=202.66, p=5.69 x 10 -70 , □2=0.91). There was also a significant interaction of time post-capsaicin by drug condition (F(7,140)=193.69, p=1.00 x 10 -70 , □2=0.91) and time post-capsaicin by sex (F(7,140)=2.31, p=0.0293, □2=0.10). There were no main effects of sex or sex by drug interactions found. At 180 DPI a four-way mixed repeated measures ANOVA comparing both male and female TBI+Ex PRZ and VEH treated rats revealed significant main effects of drug condition (F(1,20)=2546.36, p=1.46 x 10 -22 , □2=0.99), and time post-capsaicin (F(7,140)=185.55, p=1.50 x 10 -67 , □2=0.90). There was also a significant interaction of time post-capsaicin by drug condition (F(7,140)=193.27, p=1.15 x 10 -68 , □2=0.91) and time post-capsaicin by drug condition (F(7,140)=2.31, p=0.0295, □2=0.10). There were no main effects of sex or sex by drug interactions found. ⋇ = TBI+Ex+PRZ versus TBI+Ex+VEH (p < 0.0001) by Bonferroni’s posthocs. Open arrow - PGE-2; i.pl. [1 μg/50 μl]. Closed arrow - Capsaicin; i.pl. [10 μg/10 μl]. Data are presented as mean + standard error of the mean. (n = 6 rats per cohort).

Article Snippet: Prostaglandin E2 (PGE-2) [1μg/50μl], intraplantar ( i.pl. ) (14010, Cayman Chemicals, MI, USA) a principal mediator of inflammation and pain hypersensitivity.

Techniques: Injection, Saline

Journal: bioRxiv

Article Title: Actionable spatial prostanoid barriers constrain BiTE-driven adoptive T cell immunity in intact human tumors

doi: 10.64898/2026.03.26.713601

Figure Lengend Snippet: A, Basal PGE 2 quantified in NSCLC PDE supernatants (pg/mL), shown as a heatmap illustrating inter-lesion variability. B, Correlation between basal PGE 2 and MSLN-BiTE-induced CD8+ T cell activation in NSCLC PDEs, expressed as FC in CD8 + CD137 + frequencies relative to UT CTRL (Holm-corrected P value shown). C, Correlation between basal PGE 2 and MSLN-BiTE-induced granzyme B release in NSCLC PDEs, expressed as log2 FC relative to UT CTRL per patient (r and P value shown). D, COX blockade rescue experiment in two NSCLC cases (Lung 31 and Lung 33): PDEs were pretreated with ketorolac (COXi, 1 µM) for 1 h prior to co-culture with MSLN-BiTE T cells, and CD8 + CD137 + activation together with granzyme B, CXCL9, and CXCL10 were quantified as FC over UT CTRL. E, Pharmacodynamic verification of prostanoid pathway inhibition in the HGSOC validation cohort (n = 11), showing PGE 2 levels at baseline and after COXi (ketorolac, 1 µM) or EP2/EP4 inhibition (EP2/4i, 1 µM). F, CD8 + T cell activation in HGSOC PDE co-cultures, reported as %CD8 + CD137 + among CD8 + cells across conditions (condition matrix shown). G-H, FLAG-based discrimination of engineered versus bystander/resident CD8 + T cells in HGSOC PDEs, showing frequencies of CD8 + CD137 + FLAG + exogenous cells ( G ) and CD8 + CD137 + FLAG - bystander/ endogenous cells ( H ). (I) IFNγ secretion in HGSOC PDE co-cultures, expressed as FC over UT CTRL across conditions. ( J ) Cytokine and chemokine coordination across HGSOC PDEs visualized as Pearson correlation matrices for untreated, MSLN-BiTE, MSLN-BiTE + COXi, and MSLN-BiTE + EP2/4i conditions (circle size indicates correlation magnitude; color indicates direction). K, Myeloid polarization in HGSOC PDEs quantified as %CD68 + CD80 + among CD68 + macrophages across conditions. ( L-N ) TNFα ( L ), GM-CSF ( M ), and IL-10 ( N ) secretion in HGSOC PDE co-cultures, expressed as FC over UT CTRL. Boxplots show median and interquartile range with whiskers as displayed; each dot represents one patient. For HGSOC panels ( E-N ), statistics were performed on patient-level values using one-way repeated-measures ANOVA with multiple-comparisons correction as indicated (GraphPad Prism v10.1.2). Figure generated with BioRender.

Article Snippet: Basal prostaglandin E2 levels in culture supernatants from PDEs were quantified using a PGE 2 ELISA kit (Cayman Chemical, 514010) according to the manufacturer’s instructions.

Techniques: Activation Assay, Co-Culture Assay, Inhibition, Biomarker Discovery, Generated

Journal: Veterinary Sciences

Article Title: Celecoxib Inhibits Vasculogenic Mimicry and Induces Apoptosis in the D17 Canine Osteosarcoma Cell Line via the COX-2/PGE2 Signaling Axis

doi: 10.3390/vetsci13030288

Figure Lengend Snippet: Celecoxib inhibits PGE 2 production in D17 canine osteosarcoma cells in a dose-dependent manner. D17 cells were treated with celecoxib (0, 60, and 80 μM) for 48 h. PGE 2 concentrations in the culture medium were quantified by ELISA. Data represent mean ± SD. * p < 0.01 versus vehicle control by one-way ANOVA followed by Dunnett’s post hoc test.

Article Snippet: PGE 2 levels were quantified using the Parameter TM PGE 2 assay kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Veterinary Sciences

Article Title: Celecoxib Inhibits Vasculogenic Mimicry and Induces Apoptosis in the D17 Canine Osteosarcoma Cell Line via the COX-2/PGE2 Signaling Axis

doi: 10.3390/vetsci13030288

Figure Lengend Snippet: Exogenous PGE 2 rescues celecoxib-inhibited vasculogenic mimicry formation in D17 canine osteosarcoma cells. ( a ) Representative phase-contrast images of VM formation in D17 cells treated with vehicle control, 80 μM celecoxib, or 80 μM celecoxib + 100 ng/mL PGE2 at 7 h and 24 h post-seeding on Matrigel. ( b ) Quantitative analysis of VM network parameters (master junctions, master segments, meshes) at 7 h using ImageJ software. ( c ) Quantitative analysis of VM network parameters at 24 h. Magnification, ×40. Data represent mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 versus vehicle control by one-way ANOVA followed by Dunnett’s post hoc test.

Article Snippet: PGE 2 levels were quantified using the Parameter TM PGE 2 assay kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Control, Software