Journal: mBio

Article Title: Synergistic induction of PGE2 by oral pathogens and TNF promotes gingival fibroblast-driven stromal-immune cross-talk in periodontitis

doi: 10.1128/mbio.00046-25

Figure Lengend Snippet: PGE2 production and COX-2 expression are synergistically induced by oral bacteria and inflammatory cytokines in GFs. (A) PGE2 production by GFs infected with F. nucleatum ( Fn ) ( n = 5) or P. gingivalis ( Pg ) ( n = 7) with or without TNF (10 ng/mL) for 24 h. Data are shown as mean + SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; two-way ANOVA followed by Tukey multiple comparison test. (B and C) Western blot analysis of COX-2 levels in GFs infected with F. nucleatum in the absence (ctrl) or presence of IFNα (100 U/mL), IL-1β (10 pg/mL), or IL-1α (10 pg/mL) for 24 h. Actin was used as a loading control, and results representative of two independent experiments are shown.

Article Snippet: After 24 h, cell-free supernatants were collected, and the level of PGE2 was determined using Prostaglandin E2 Express ELISA Kit (Cayman Chemical).

Techniques: Expressing, Bacteria, Infection, Comparison, Western Blot, Control

Journal: mBio

Article Title: Synergistic induction of PGE2 by oral pathogens and TNF promotes gingival fibroblast-driven stromal-immune cross-talk in periodontitis

doi: 10.1128/mbio.00046-25

Figure Lengend Snippet: p38 MAPK activation drives synergistic induction of COX-2 and PGE2 by oral bacteria and TNF. (A) Western blot analysis of COX-2 in GFs pretreated with DMSO or MAPK inhibitors (p38i, JNKi, and ERKi) prior to infection with F. nucleatum ( Fn ) in the absence (ctrl) or presence of 10 ng/mL TNF for 24 h. Representative blots (upper panel) and results of densitometric analysis for selected conditions ( n = 3, bottom panel) are shown. * P < 0.05; ratio paired t test. (B) Western blot analysis of COX-2 in GFs pretreated with DMSO or p38 inhibitor (p38i) prior to infection with P. gingivalis ( Pg ) without (ctrl) or with stimulation with 10 ng/mL TNF for 24 h. Representative blots (upper panel) and results of densitometric analysis ( n = 3, bottom panel) are shown. * P < 0.05; One-way ANOVA followed by Bonferroni multiple comparison test. (C) PGE2 production by GFs ( n = 5) pretreated with DMSO or p38i prior to infection with P. gingivalis or F. nucleatum in the absence (ctrl) or presence of 10 ng/mL TNF for 24 h. Data are presented as mean + SEM. * P < 0.05; ** P < 0.01; ratio paired t test.

Article Snippet: After 24 h, cell-free supernatants were collected, and the level of PGE2 was determined using Prostaglandin E2 Express ELISA Kit (Cayman Chemical).

Techniques: Activation Assay, Bacteria, Western Blot, Infection, Comparison

Journal: mBio

Article Title: Synergistic induction of PGE2 by oral pathogens and TNF promotes gingival fibroblast-driven stromal-immune cross-talk in periodontitis

doi: 10.1128/mbio.00046-25

Figure Lengend Snippet: GF-derived PGE2 promotes IL8 expression in MDMs. (A) Schematic representation of the experimental setup. Created in BioRender. Grabiec, A. (2025) https://BioRender.com/k40a236 . (B) qPCR analysis of IL8 expression in MDMs stimulated with PGE2 in the absence (ctrl) or presence of 10 ng/mL TNF for 24 h. ** P < 0.01; *** P < 0.001; Two-way ANOVA followed by Tukey multiple comparison test. (C and D) Western blot analysis of COX-2 in GFs transfected with control non-targeting siRNA (si-ctrl) or COX-2-targeting siRNA (si-COX-2) before infection with F. nucleatum ( Fn ) without (ctrl) or with 10 ng/mL TNF for 24 h. Representative blots (upper panel) and results of densitometric analysis for selected conditions ( n = 3, bottom panel) are shown. * P < 0.05; ratio paired t test. (E) qPCR analysis of IL8 expression in MDMs after 24 h stimulation with TNF (50 ng/mL) in the presence of conditioned media from GFs ( n = 4) that were treated as in (C). Data are presented as mean + SEM. * P < 0.05; *** P < 0.001; comparisons between MDMs subjected to GF conditioned media from different transfection conditions were performed using ratio paired t test, whereas comparisons between MDM responses to GF conditioned media from different infection/stimulation conditions were performed using one-way ANOVA followed by Bonferroni multiple comparison test.

Article Snippet: After 24 h, cell-free supernatants were collected, and the level of PGE2 was determined using Prostaglandin E2 Express ELISA Kit (Cayman Chemical).

Techniques: Derivative Assay, Expressing, Comparison, Western Blot, Transfection, Control, Infection

Journal: International immunopharmacology

Article Title: Interleukin 1β and Prostaglandin E2 affect expression of DNA methylating and demethylating enzymes in human gingival fibroblasts.

doi: 10.1016/j.intimp.2019.105920

Figure Lengend Snippet: Fig. 1. IL-1β and PGE2 affect expression of DNMT1, DNMT3a and TET1. (A and C) Total RNA was isolated from untreated HGF cells (control) or cells treated for 24 h with IL-1β or PGE2 at the indicated doses. (B and D) RNA was isolated at the indicated times after addition of IL-1β (10 ng/ml, 0.57 μM) or PGE2 (100 μM). mRNA levels of DNMT1, DNMT3a and TET1 were determined by real-time PCR. (E) Whole cell lysates were isolated from HGF cultures that were untreated (control) or treated for 24 h with IL-1β (10 ng/ml, 0.57 μM) or PGE2 (100 μM). DNMT1 and DNMT3a protein levels were determined by ELISA. Graphs show mRNA or protein levels expressed relative to untreated controls as box and whisker plots showing interquartile ranges, with the mean marked by X. At least three different HGF cell lines derived from different donors were used in each experiment. Statistical significance (*p < 0.05, **p < 0.01) vs untreated control was determined by One-way ANOVA followed by Dunnett’s test for multiple comparisons.

Article Snippet: PGE2 levels in conditioned media from untreated control cultures or cultures treated with IL-1β (10 ng/ml, 0.57 μM) and/or NS398 (0.1 μM) for 24 h were determined using the Prostaglandin E2 Parameter Assay Kit (R&D Systems) according to the manufacturer’s suggested protocol.

Techniques: Expressing, Isolation, Control, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Whisker Assay, Derivative Assay

Journal: International immunopharmacology

Article Title: Interleukin 1β and Prostaglandin E2 affect expression of DNA methylating and demethylating enzymes in human gingival fibroblasts.

doi: 10.1016/j.intimp.2019.105920

Figure Lengend Snippet: Fig. 2. IL-1β and PGE2 affect global levels of 5mC and 5hmC. Global levels of 5mC (A) and 5hmC (B) were determined by MethylFlash ELISA-like assays using DNA isolated from HGF that were untreated (control) or treated with IL- 1β (10 ng/ml, 0.57 μM) or PGE2 (100 μM) for 24, 48 or 72 h. Graphs show percent 5mC or 5hmC expressed relative to untreated controls, as box and whisker plots showing interquartile ranges, with the mean marked by X. Five different HGF cell lines derived from different donors were used. Statistical significance (*p < 0.05, **p < 0.01) vs untreated control was determined by One-way ANOVA followed by Dunnett’s test for multiple comparisons. Global levels of 5hmC were lower as compared to 5mC, but in the range expected based on other studies [75,79].

Article Snippet: PGE2 levels in conditioned media from untreated control cultures or cultures treated with IL-1β (10 ng/ml, 0.57 μM) and/or NS398 (0.1 μM) for 24 h were determined using the Prostaglandin E2 Parameter Assay Kit (R&D Systems) according to the manufacturer’s suggested protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Control, Whisker Assay, Derivative Assay

Journal: International immunopharmacology

Article Title: Interleukin 1β and Prostaglandin E2 affect expression of DNA methylating and demethylating enzymes in human gingival fibroblasts.

doi: 10.1016/j.intimp.2019.105920

Figure Lengend Snippet: Fig. 3. Effects of IL-1β and PGE2 on mRNA expression of genes that showed changes in methylation at specific CpG in their promoters. Total RNA was isolated from HGF cultures that were untreated (control) or treated with IL-1β (10 ng/ml, 0.57 μM) or PGE2 (100 μM) for 24 or 48 h. mRNA levels of IL-8 (A and B) and CD40 and TNFRSF10C (C and D) were determined by real-time PCR and expressed relative to levels in untreated controls. Graphs show data from three different HGF cell cultures derived from three different donors, with mRNA levels expressed relative to untreated controls and depicted in box and whisker plots showing interquartile ranges, with the mean marked by X. Statistical significance (*p < 0.05, **p < 0.01) vs untreated control was determined by One-way ANOVA followed by Dunnett’s test for multiple comparisons.

Article Snippet: PGE2 levels in conditioned media from untreated control cultures or cultures treated with IL-1β (10 ng/ml, 0.57 μM) and/or NS398 (0.1 μM) for 24 h were determined using the Prostaglandin E2 Parameter Assay Kit (R&D Systems) according to the manufacturer’s suggested protocol.

Techniques: Expressing, Methylation, Isolation, Control, Real-time Polymerase Chain Reaction, Derivative Assay, Whisker Assay

Journal: International immunopharmacology

Article Title: Interleukin 1β and Prostaglandin E2 affect expression of DNA methylating and demethylating enzymes in human gingival fibroblasts.

doi: 10.1016/j.intimp.2019.105920

Figure Lengend Snippet: Fig. 4. IL-1β effects on gene expression are not altered by inhibition of COX-2 mediated PGE2 production. HGF cell cultures were untreated (control) or pre- treated with NS398 (0.1 μM) for 1 h before addition of IL-1β (10 ng/ml, 0.57 μM). Conditioned media and total RNA were harvested 24 h after addition of IL-1β. (A) PGE2 levels in conditioned media were determined by Prostaglandin E2 Parameter Assay Kit (R&D Systems). (B) DNMT1, DNMT3a and TET1 mRNA levels were determined by real-time PCR and expressed re- lative to untreated control. Graphs show data from three to five different HGF cell lines derived from different donors, expressed relative to untreated con- trols, as box and whisker plots showing interquartile ranges, with the mean marked by X. Statistical significance (**p < 0.01 vs untreated control, $p < 0.05 vs IL-1β alone)) was determined by Two-way ANOVA followed by Tukey’s test for multiple comparisons. There was no statistical significance detected in comparison of PGE2 levels in IL-1 + NS398 vs control or vs Ns398 alone; and also no significant difference in mRNA levels between IL and 1+ NS389 vs IL-1 alone.

Article Snippet: PGE2 levels in conditioned media from untreated control cultures or cultures treated with IL-1β (10 ng/ml, 0.57 μM) and/or NS398 (0.1 μM) for 24 h were determined using the Prostaglandin E2 Parameter Assay Kit (R&D Systems) according to the manufacturer’s suggested protocol.

Techniques: Gene Expression, Inhibition, Control, Real-time Polymerase Chain Reaction, Derivative Assay, Whisker Assay, Comparison

Journal: International immunopharmacology

Article Title: Interleukin 1β and Prostaglandin E2 affect expression of DNA methylating and demethylating enzymes in human gingival fibroblasts.

doi: 10.1016/j.intimp.2019.105920

Figure Lengend Snippet: Fig. 6. An EP4 receptor antagonist blocks the effect of PGE2 on expression of DNMT3a. HGF cultures were pre-treated with EP4 receptor antagonist ONOAE3208 (0.01 μM) for one hour before addition of IL-1β (10 ng/ml, 0.57 μM) or PGE2 (100 μM) for 24 h. Levels of DNMT3a mRNA were de- termined by real-time PCR and expressed relative to untreated controls. Four different HGF cell lines derived from different donors were used. The graph shows data in box and whisker plots showing interquartile ranges, with the mean marked by X. Statistical significance (**p < 0.01 vs untreated control, $ $p < 0.01 vs PGE2 alone) was determined by Two-way ANOVA with post-hoc Tukey. No statistical significance was detected between IL-1+antagonist vs IL-1 alone, or between control vs PGE2+antagonist.

Article Snippet: PGE2 levels in conditioned media from untreated control cultures or cultures treated with IL-1β (10 ng/ml, 0.57 μM) and/or NS398 (0.1 μM) for 24 h were determined using the Prostaglandin E2 Parameter Assay Kit (R&D Systems) according to the manufacturer’s suggested protocol.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Derivative Assay, Whisker Assay, Control

Journal: Anatomy Journal of Africa

Article Title: The histochemical and biochemical effects of coconutmilk in Wistar rats with ethanol-induced gastric ulcer

doi: 10.4314/aja.v8i2.188223

Figure Lengend Snippet: Figure 6: Prostaglandin E2 Concentration in the stomach of coconut milk-treated rats following ethanol- induced gastric ulcer. Each bar represents Mean ± S.E.M. in each group. a, b, bc, c within column signifies that means with different letters differs significantly at p < 0.001 while means with the same letters does not differ significantly at p < 0.001.

Article Snippet: After the stomach has been harvested, opened along the greater curvature and washed in normal saline, it was placed on ice and the mucosa was gently scraped into a specimen bottle and kept in a freezer at -20°C.Nitric oxide levels in the gastric mucosa were measured as total nitrite and nitrate level with the use of Greiss reagent by the method of Moshage and coworkers (Moshage et al., 1995).Tissue concentration of Prostaglandin E2 was determined by using prostaglandin E2 ELISA Assay kit (Elabscience laboratory, China).

Techniques: Concentration Assay