Journal: Veterinary Sciences

Article Title: CD4 Molecule Plays an Important Role in the Inflammatory Response Induced by Japanese Encephalitis Virus Infection

doi: 10.3390/vetsci13030254

Figure Lengend Snippet: CD4 Knockdown Reduces p-STAT1 Protein Levels in TM3 Cells Post-JEV Infection. Note: *** indicates p < 0.001 , representing a statistically significant difference compared to the control group. ( A ) TM3 and CD4.KD cells were infected with JEV (MOI = 1) for 48 h, and cell samples were collected for Western blot analysis to assess p-STAT1 protein levels. ( B ) To quantify the expression level of the p-STAT1 protein, gray value analysis of the bands from three biological replicates was performed using ImageJ software. Statistical analysis revealed a significant difference in p-STAT1 protein expression between CD4.KD and TM3 cells ( p = 0.0006, p < 0.01) (the original pictures can be found in ).

Article Snippet: The STAT1 agonist RO8191 (dissolved in DMSO) was purchased from MedChemExpress (MCE, Monmouth Junction, NJ, USA) and diluted to the working concentration in serum-free DMEM prior to use.

Techniques: Knockdown, Infection, Control, Western Blot, Expressing, Software

Journal: Veterinary Sciences

Article Title: CD4 Molecule Plays an Important Role in the Inflammatory Response Induced by Japanese Encephalitis Virus Infection

doi: 10.3390/vetsci13030254

Figure Lengend Snippet: CD4 Gene Knockdown Significantly Inhibits the Activating Effect of RO8191 on the JAK-STAT1 inflammatory pathway, and This Inhibitory Phenotype Can Be Reversed by CD4 Replenishment. Note: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.0001, all representing significant differences compared with the control group. ( A ) Three models were established: normal TM3 cells, CD4.KD cells, and CD4 replenished cells. Each group was divided into a control group and an RO8191 pre-treatment experimental group. After inoculating with JEV at an MOI of 1 and culturing for 48 h, Western blot analysis was performed to detect the expression levels of p-STAT1 protein. ( B ) Gray value analysis of the bands from three biological replicates was performed using ImageJ to quantify the p-STAT1 protein (the original pictures can be found in ).

Article Snippet: The STAT1 agonist RO8191 (dissolved in DMSO) was purchased from MedChemExpress (MCE, Monmouth Junction, NJ, USA) and diluted to the working concentration in serum-free DMEM prior to use.

Techniques: Knockdown, Control, Western Blot, Expressing

Journal: Veterinary Sciences

Article Title: CD4 Molecule Plays an Important Role in the Inflammatory Response Induced by Japanese Encephalitis Virus Infection

doi: 10.3390/vetsci13030254

Figure Lengend Snippet: CD4 Replenishment Reverses the Inhibitory Effect of CD4 Knockdown on the Production of Inflammatory Factors (IL-1β, IL-6, IFN-γ, TNF-α, IL-10) Induced by JEV Infection in TM3 Cells. Note: * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.0001, all representing significant differences compared with the control group. Three models were established: normal TM3 cells, CD4.KD cells, and CD4 replenished cells. Each group was divided into a control group and an RO8191 pre-treatment experimental group. After inoculating with JEV at an MOI of 1 and culturing for 48 h: ( A ) Western blot analysis was performed to detect the expression levels of IL-1β protein. ( B ) Gray value analysis of the bands from three biological replicates was performed using ImageJ to quantify the IL-1β protein. ( C ) The expression levels of IL6 mRNA in each group of cells were detected by RT-qPCR. ( D ) The expression levels of IFN-γ mRNA in each group of cells were detected by RT-qPCR. ( E ) The expression levels of TNF-α mRNA in each group of cells were detected by RT-qPCR. ( F ) The expression levels of IL-10 mRNA in each group of cells were detected by RT-qPCR (the original pictures can be found in ).

Article Snippet: The STAT1 agonist RO8191 (dissolved in DMSO) was purchased from MedChemExpress (MCE, Monmouth Junction, NJ, USA) and diluted to the working concentration in serum-free DMEM prior to use.

Techniques: Knockdown, Infection, Control, Western Blot, Expressing, Quantitative RT-PCR

Journal: bioRxiv

Article Title: RO8191, a new compound for initiating embryo implantation in mice

doi: 10.1101/2024.09.30.615945

Figure Lengend Snippet: (A) The experimental procedure of the artificial DI mouse. Ovariectomy (OVX) and subcutaneous administration of siliconized medroxyprogesterone acetate (MPA) were performed on D3. A single injection of oil or RO8191 was performed on D7. Females were euthanized and dissected on D10. (B) Representative images of the gross uterine morphology. Blastocysts were recovered from oil-treated mice (white arrowheads). Implantation sites (yellow arrowheads on right panel) were visible in DI mice after RO8191 treatment. Scale bars: 1 cm for uterine images and 200 μm for blastocysts. (C, D) Implantation rate (C) and the number of implantation sites (D) in the oil and RO8191-treated DI mice. * indicates significantly different (p < 0.0001). (E) Representative histological image of uterine cross-sections of implantation sites on D10 after RO8191 treatment in DI mice. Right panels are higher magnification of left panels indicated by yellow line. Lower panels indicated abnormal embryo development and decidual reaction. Scale bars: 100 μm. dec: decidua; em: embryo.

Article Snippet: The RO8191 (T22142, TargetMol) was dissolved in sesame oil (196-15385, FujiFilm Wako Pure Chemicals), and a single intraperitoneal (i.p.) injection of RO8191 (400 μg/head; dissolved in sesame oil) or sesame oil was performed at 1300 h on D7.

Techniques: Injection

Journal: bioRxiv

Article Title: RO8191, a new compound for initiating embryo implantation in mice

doi: 10.1101/2024.09.30.615945

Figure Lengend Snippet: (A) The experimental procedure of the genetically modified mice. For control, a single oil injection was performed on D4. For cKOs, a single injection of oil or RO8191 was performed on D4. Females were euthanized and dissected on D7. (B-D) Uterine morphology of cKOs mice after the injection of oil or RO8191. The white arrowheads indicate the presumptive implantation site. Scale bars: 1 cm. (E) The implantation rate in cKOs mice after oil or RO8191 administration. (F) The number of implantation sites in cKOs mice after the injection of oil or RO8191. (G) The size of implantation sites in cKOs mice after oil or RO8191 administration. * indicates significantly different (p < 0.05).

Article Snippet: The RO8191 (T22142, TargetMol) was dissolved in sesame oil (196-15385, FujiFilm Wako Pure Chemicals), and a single intraperitoneal (i.p.) injection of RO8191 (400 μg/head; dissolved in sesame oil) or sesame oil was performed at 1300 h on D7.

Techniques: Genetically Modified, Control, Injection

Journal: bioRxiv

Article Title: RO8191, a new compound for initiating embryo implantation in mice

doi: 10.1101/2024.09.30.615945

Figure Lengend Snippet: (A, B) Control ( Lifr f/f ) uterus after sesame oil treatment. (C, D) Stat3 cKO, (E, F) Gp130 cKO, and (G, H) Lifr cKO uterus after RO8191 treatment. Higher magnification in yellow frames inserted in D and F. Scale bar, 100 µm. dec: decidua; em: embryo; bv: blood vessels; le: luminal epithelium; li: leukocyte infiltration.

Article Snippet: The RO8191 (T22142, TargetMol) was dissolved in sesame oil (196-15385, FujiFilm Wako Pure Chemicals), and a single intraperitoneal (i.p.) injection of RO8191 (400 μg/head; dissolved in sesame oil) or sesame oil was performed at 1300 h on D7.

Techniques: Control

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