Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: RO8191 activity on CHKV -nanoluc replication. A) Representative structure of CHIKV- nanoluc genome [PDB: 6NK5]( Basore et al., 2019 ; Metz and Pijlman, 2016 ). B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ). C) Representative scheme of the infection assays. BHK-21 cells were treated with two-fold serial dilutions of RO8191 at concentrations ranging from 0.46 to 60 µM. CHIKV- nanoluc replication was quantified by measuring nanoluciferase activity (indicated by a black square) and cell viability using MTT assay (indicated by a gray circle). Mean ± SD values from a minimum of three independent experiments, each measured in triplicate, are represented. All images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Activity Assay, Infection, MTT Assay, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: RO8191 activity on different stages of CHIKV- nanoluc replicative cycle in BHK-21 cells. A) BHK-21 cells were treated with RO8191 for 1 h, cells were washed with PBS to remove the compound, and were infected with CHIKV- nanoluc at MOI 0.1 for 1 h. Then, the medium was removed, cells were washed to remove unbound virus, and fresh medium was added. B) BHK-21 cells were infected with CHIKV- nanoluc (MOI 0.1) and simultaneously treated with RO8191 for 1 h. Then, cells were washed to completely remove the inoculum, and a fresh medium was added. C) RO8191 and CHIKV- nanoluc at MOI 5 were incubated for 1 h (representative inoculum). Then, the inoculum was added to the cells for an additional hour. Cells were washed to remove the inoculum and fresh medium was added. D) BHK-21 cells were infected with CHIKV- nanoluc (MOI 0.1) for 1 h, cells were washed with PBS to remove unbound virus, and were treated with RO8191. All infection assays were quantified 16 h.p.i through the measurement of luminescence levels. Schematic representation of each time-based assay as indicated by BHK-21 cells (black bars), RO8191 (gray bars), CHIKV- nanoluc (orange bars), and CHIKV-RO8191 inoculum (blue tube). E) Schematic representation of the time-based BHK-CHIKV cell assay as indicated by the BHK-CHIKV-NCT cells (black bars) and RO8191 (gray bars). BHK-CHIKV-NCT cells were incubated for 72 h with the RO8191 compound at concentrations ranging from 60 to 0.46 µM. Then, cells were washed with PBS and lysed with Passive Lysis Buffer to measure the activity of Renilla . The reduction of Renilla activity in BHK-CHIKV-NCT cells is shown by 50% of the highest concentrations (EC 50 ) of RO8191 (black squares) and an effect in cell viability (CC 50 ) using MTT assay (gray circles). Mean ± SD values of a minimum of three independent experiments, each measured in triplicate. (***) P <0.001, (**) P <0.01, and (*) P <0.05 are represented. Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Activity Assay, Infection, Virus, Incubation, Lysis, MTT Assay, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: BHK-21 cells express type-1 IFN (α/β) during RO8191 treatment. A) Percentage of CHIKV WT infection by measuring mRNA-CHIKV WT in supernatant of BHK-21 cells. B) CHIKV WT RNA titers by measuring mRNA-CHIKV WT in supernatant of BHK-21 cells. C) Percentage of CHIKV WT infection by measuring mRNA-CHIKV WT per mL in cell lysates of BHK-21 cells. D) CHIKV WT RNA titers by measuring mRNA-CHIKV WT per mL detected in cell lysates of BHK-21 cells. The CHIKV infection is presented in black bars while CHIKV in the presence of RO8191 treatment is demonstrated in gray bars. E) mRNA means of IFN-α expressed by BHK-21 during infection and/or treatment with RO8191. F) mRNA mean of IFN-β expressed by BHK-21 during infection and/or treatment with RO8191. BHK-21 cells were infected with CHIKV WT (MOI 0.1 PFU/well) in the presence or absence of RO8191 at 5 µM for 16 h. The supernatant and cell lysates were collected and processed to RNA extraction. cDNA was produced and mRNA levels was quantified by RT-qPCR. Mean values of three independent experiment each measured in triplicate, are represented. (****) P <0.0001. Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Infection, RNA Extraction, Produced, Quantitative RT-PCR, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: RO8191 activity against CHIKV- nanoluc infection in Vero-E6 cells. A) Representative scheme of the infection assays. Vero-E6 cells were treated with three-fold serial dilutions of RO8191 at concentrations ranging from 0.02 to 60 µM. CHIKV replication was quantified by measuring nanoluciferase activity (indicated by a black square) and cell viability using an MTT assay (indicated by a gray circle). Values of EC 50 , CC 50, and SI were obtained from the treatment of Vero-E6 cells treated with RO8191. B) Vero-E6 cells were treated with RO8191 for 1 h, then the cells were washed with PBS and infected with CHIKV- nanoluc virus at MOI 0.1 for 1 h. The medium was removed, cells were washed with PBS and fresh medium was added. C) Vero-E6 cells were infected with CHIKV- nanoluc (MOI 0.1) and simultaneously treated with RO8191 for 1 h. Then, cells were washed, and fresh medium was added. D) RO891 and CHIKV- nanoluc at MOI 5 were incubated for 1 h, then the inoculum was added to cells for an extra 1 h, cells were washed, and fresh medium was added. E) Vero-E6 cells were infected with CHIKV- nanoluc (MOI 0.1) for 1 h, cells were washed with PBS and treated with RO8191. Schematic representation of each time-based assay as indicated by Vero-E6 cells (blue bars), RO8191 (gray bars), CHIKV- nanoluc (orange bars), and CHIKV- nanoluc /RO8191 inoculum (blue tube). Mean ± SD values of a minimum of three independent experiments, each measured in triplicate, are represented. (****) P <0.0001, (***) P <0.001 and (*) P <0.05. Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Activity Assay, Infection, MTT Assay, Virus, Incubation, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: Localization and ChemPLP values showed between RO8191 and CHIKV proteins by molecular docking calculations.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques:

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: 2D and 3D interactions between RO8191 and CHIKV proteins showed by molecular docking analysis. A) RO8181 interacts with CHIKV glycoproteins mainly through Hydrogen Bond (green lines) and alkyl ligands (pink lines). B) RO8191 showed interactions with nsP1 protein mainly through Conventional Hydrogen Bonds (green circles) interactions. C) RO8191 interacts with nsP2 mainly through Alkyl (pink lines) and Hydrogen Bonds (green lines) interactions. D) RO8191 interacts with nsP3 mainly through Hydrogens Bonds ligands and interactions (green lines and circles). E) RO8191 interactions with nsP4 mainly through Conventional Hydrogens Bonds (green lines) and Halogen (blue lines) ligands. RO8191 (PDB: 3R0L) and the CHIKV proteins (glycoprotein complex [PDBid: 3N42], nsP1 [PDBid: 7DOP], nsP2 [PDBid:4ZTB], nsP3 [PDBid:6W8Z], and database representative sequence of the nsP4 extracted from the virus polyprotein [uniport-id: Q8JUX6]) were docked employing the GOLD program site using a genetic algorithm (GA) and the score ChemPL. Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Sequencing, Virus, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: Infrared spectroscopy indicates molecular interactions between CHIKV virions with RO8191. A) Representative scheme of the ATR-FTIR technology with CHIKV virions (black line), RO8191 (blue line), and CHIKV virion plus incubation with RO8191 (red line). B) The representative infrared average spectrum of normalized mean spectra of CHIKV virion (black line), RO8191 (blue line), and CHIKV virion plus RO8191 (red line) employing ATR-FTIR platform from 1800 to 800 cm −1 . C-F) The representative mean spectra of second derivative analysis to the vibrational modes at 1496 cm −1 ( C ), 1465 cm −1 ( D ), 1448 cm −1 ( E ), 1333 cm −1 ( F ), and 1078 cm −1 ( G ). Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Spectroscopy, Incubation, Generated

Journal: Discover. Oncology

Article Title: Ritanserin suppresses acute myeloid leukemia by inhibiting DGKα to downregulate phospholipase D and the Jak-Stat/MAPK pathway

doi: 10.1007/s12672-023-00737-9

Figure Lengend Snippet: Agonists of Jak-Stat/MAPK signaling rescued the ritanserin-mediated anticancer effect and activates underlying pathways. The histograms of cell viability and apoptosis cell ratio after ritanserin combined with A the MAPK, MEK/ERK agonist C16-PAF or B the JAK/STAT agonists RO8191 for 48 h. Data are presented as the mean ± S.D. of triplicate experiments. (*p < 0.05; ***p < 0.001.) C The protein expression levels of p-MEK1/2(Ser217/221), MEK1/2, p-ERK1/2(Thr202/Tyr204) and ERK1/2 were assessed after treatment with ritanserin, C16-PAF or both for 48 h. D The protein expression levels of JAK1, JAK3, p-STAT5(Typ694), STAT5, p-STAT3(Tyr705) and STAT3 were assessed after treatment with ritanserin, RO8191 or both for 48 h

Article Snippet: The protein agonists applied were as follows: SphK1 agonist K6PC-5(HY-124042, MedChemExpress); MEK/ERK agonists C16-PAF (HY-108635, MedChemExpress); JAK/STAT agonists RO8191 (T22142, TargetMol).

Techniques: Expressing

Journal: Cell Death Discovery

Article Title: EZH2–STAT3 signaling pathway regulates GSDMD-mediated pyroptosis in glioblastoma

doi: 10.1038/s41420-024-02105-0

Figure Lengend Snippet: A , B Immunoblotting shows protein levels of NLRP3, GSDMD, N-GSDMD in control, SH-4–54, and SH-4–54 + RO8191 groups after SH-4–54 treatment. C Microscopic observation of the morphological effects of RO8191 on U87 and LN229 cells after SH-4–54 treatment. D , E EdU assay demonstrates proliferation capabilities in control, SH-4–54, and SH-4–54 + RO8191 groups. For A , B , data are presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, N.S. not significant by ANOVA.

Article Snippet: Inhibitors and agonists were purchased from MedChemExpress (New Jersey, USA) - DZNep and RO8191, while SH-4–54 was purchased from Selleck (Houston, Texas, USA).

Techniques: Western Blot, Control, EdU Assay

Journal: Molecular Medicine Reports

Article Title: Fucoxanthin alleviates chondrocyte inflammation and osteoarthritis by modulating the JAK2/STAT3 signaling pathway

doi: 10.3892/mmr.2026.13933

Figure Lengend Snippet: Effects of FUC and RO8191 on cartilage degeneration and JAK2/STAT3 signaling in vivo . Representative (A) H&E-stained full-joint coronal sections and (B) corresponding high-magnification images of articular cartilage from ACLT rats treated with 200 mg/kg FUC, with or without RO8191 (n=6). Representative (C) Safranin O-Fast Green-stained full-joint coronal sections and (D) corresponding high-magnification images showing cartilage proteoglycan content (n=6). (E) OARSI scores of full-joint coronal sections, with corresponding high-magnification views (n=6). (F) Immunohistochemical staining and quantitative analysis of p-JAK2, total JAK2, p-STAT3 and total STAT3 in articular cartilage (n=6). Scale barrs: 200 µm for (A,C) full-joint coronal sections; 100 µm for (B,D) high-magnification sections. Data are presented as the mean ± SD. **P<0.01. ACLT, anterior cruciate ligament transection; FUC, fucoxanthin; OARSI, Osteoarthritis Research Society International; p, phosphorylated; JAK, Janus kinase.

Article Snippet: For RO8191 treatment, chondrocytes were co-incubated with 5 μM RO8191 together with IL-1β, FUC or both at 37°C in 5% CO2 for 24 h. Recombinant rat IL-1β (cat. no. RP01788; ABclonal Biotech Co., Ltd.) was used for cell stimulation at a concentration of 10 ng/ml at 37°C for 24 h. For control groups, untreated chondrocytes served as the negative control and IL-1β-only treated chondrocytes served as the model control.

Techniques: In Vivo, Staining, Immunohistochemical staining

Journal: Molecular Medicine Reports

Article Title: Fucoxanthin alleviates chondrocyte inflammation and osteoarthritis by modulating the JAK2/STAT3 signaling pathway

doi: 10.3892/mmr.2026.13933

Figure Lengend Snippet: FUC reduces inflammatory responses associated with JAK2/STAT3 signaling in vitro . Primary rat chondrocytes were stimulated with IL-1β (10 ng/ml, 24 h) and treated with FUC (15 µM) and/or the JAK2 activator RO8191. (A) Western blot analysis and semi-quantitative evaluation of JAK2/STAT3 signaling components and inflammatory/catabolic protein expression (n=3). (B) Reverse transcription-quantitative PCR analysis of IL-6, MMP3, MMP13 and ADAMTS5 (n=3). (C) ELISA analysis of IL-6 secretion. (D) Immunofluorescence staining of MMP3 in chondrocytes with nuclei counterstained with DAPI, and corresponding quantitative fluorescence intensity analysis. Scale bar, 50 µm. Data are presented as the mean ± SD. *P<0.05; **P<0.01. NC, negative control; FUC, fucoxanthin; ADAMTS, a disintegrin and metalloproteinase with thrombospondin motifs; iNOS, inducible nitric oxide synthase; p, phosphorylated; JAK, Janus kinase.

Article Snippet: For RO8191 treatment, chondrocytes were co-incubated with 5 μM RO8191 together with IL-1β, FUC or both at 37°C in 5% CO2 for 24 h. Recombinant rat IL-1β (cat. no. RP01788; ABclonal Biotech Co., Ltd.) was used for cell stimulation at a concentration of 10 ng/ml at 37°C for 24 h. For control groups, untreated chondrocytes served as the negative control and IL-1β-only treated chondrocytes served as the model control.

Techniques: In Vitro, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Fluorescence, Negative Control

Journal: Virology Journal

Article Title: Reovirus resistance in tumors mediated by elevated ISG expression: overcoming therapeutic resistance via JAK/STAT pathway modulation

doi: 10.1186/s12985-026-03118-0

Figure Lengend Snippet: The Impact of JAK-STAT Signaling Pathway-Induced ISGs on Reovirus Replication and Oncolytic Activity. A Relative expression of OAS1/2/3 , ISG15 , and MX2 mRNA in HS and LS cells measured by RT-qPCR. B COC1/DDP cells were pretreated with RO8191 for 24 h, followed by treatment with 5 MOI reovirus for an additional 24 h. Subsequently, the relative expression of OAS1/2/3 , ISG15 , and MX2 mRNA in COC1/DDP cells was measured using RT-qPCR. C Quantification of viral RNA levels by RT-qPCR. SKOV3 and COC1/DDP cells were treated as described in ( B ). D Assessment of cell viability using the CCK-8 assay. SKOV3 and COC1/DDP cells were pretreated with RO8191 for 24 h, followed by treatment with 5 MOI reovirus for 48 h. Statistical analyses were performed using the Student’s t -test ( n = 3 per group; * p < 0.05, ** p < 0.01, *** p < 0.001). Data are presented as mean ± SD

Article Snippet: SKOV3 and COC1/DDP cells were pre-treated with 3 μM RO8191 (MCE, USA) or 10 μM IFN-alpha-IFNAR-IN-1 (MCE, USA) for 24 h, followed by exposure to 5 MOI reovirus for 48 h [ , ].

Techniques: Activity Assay, Expressing, Quantitative RT-PCR, CCK-8 Assay

Journal: Advanced Science

Article Title: hESC‐Derived Epicardial Cells Promote Repair of Infarcted Hearts in Mouse and Swine

doi: 10.1002/advs.202300470

Figure Lengend Snippet: The IFN‐I receptor agonist RO8191 attenuates the anti‐inflammatory and cardiac reparative effects of hEPs. A) Cardiac performance measured by echocardiography. All mice received intragastric administration of 10 mg kg −1 RO8191 or vehicle control (corn oil) 30 min before MI surgery. n = 6, 8, 6, and 8 in the MI, MI + hEPs, MI + RO8191, and MI + hEPs + RO8191 groups. ** p < 0.01, *** p < 0.001, and n.s.(no significant difference) versus the MI group. # p < 0.05, ## p < 0.01, and n.s. as indicated. B) Representative images and quantitative analysis of scar sizes stained with Masson's trichrome at day 28 post‐MI. n = 6 to 7. Scale bar: 1 mm. C,D) Representative images and quantitative data of immunofluorescent staining for CD31 + endothelial cells (C, n = 5 to 6) and LYVE1 + lymphatic vessels (D, n = 4 to 7) in the border zones at day 28 post‐MI. Scale bar: 50 µm. E) Representative images and quantitative data of immunofluorescent staining for TUNEL + CMs in the border zones at day 3 post‐MI. n = 4 to 5. Scale bar: 20 µm. F) Representative images and quantitative data of immunofluorescent staining for CD68 + cells and CD68 + CD206 + macrophages in the border zones at day 3 post‐MI. n = 5 to 6. Scale bar: 50 µm. Data are means ± S.E.M. Two‐way ANOVA followed by Bonferroni's multiple analysis in A to F. * p < 0.05, ** p < 0.01, and *** p < 0.001. n.s.: no significant difference.

Article Snippet: RO8191 (APExBIO) was diluted with corn oil and intragastrically administrated at 10 mg kg −1 within 30 min before MI injury.

Techniques: Control, Staining, TUNEL Assay