pge2 elisa pge2 levels  (R&D Systems)

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NitraTh epitope-based neoantigen vaccines for effective tumor immunotherapy

doi: 10.1007/s00262-024-03830-2

Figure Lengend Snippet: NitraTh epitope elevates gene expression of 15pgdh accompanied by a decrease in PGE2 levels. (A) Top 10 significantly differentially enriched pathways in samples belonging to the PADRE and NitraTh clusters identified by GSEA of Hallmark gene sets. Count, number of genes contributing to the enrichment score. (B) The enrichment plot of the arachidonic acid metabolism genes shows significant enrichment in NitraTh versus PADRE in murine splenocytes on day 21 after treatment. (C-D) Elisa of PGE2 secretion from H22 tumor-bearing mouse serum (C) or tumor (D) on day 19. (E–F) Elisa of PGE2 secretion from PDX-HIS tumor-bearing mouse serum (E) or tumor (F) on day 21. (G-H) Elisa of PGE2 secretion from MC38 tumor-bearing mouse serum (G) or tumor (H) on day 23. (I) The heat map represents the abundance of the arachidonic acid metabolism genes in NitraTh versus PADRE in murine splenocytes on day 21 after treatment. (J) Slco2a1 RNA-seq expression data. TPM, transcripts per million. (K) 15pgdh expression in the splenocytes, as measured by qPCR. (L-M) 15pgdh (L) or Slco2a1 (M) expression in RAW264.7 cells as measured by QPCR. (N–O) Gene expression of 15pgdh (N) or Slco2a1 (O) in RAW264.7 cells at various drug concentrations. (P) PGE2 levels in cell culture supernatant of RAW264.7 cells. (Q) PGE2 levels in cell culture supernatant of RAW264.7 cells when using SW033291 inhibitors. (R-S) Gene expression of 15pgdh (R) or Slco2a1 (S) in BMDMs at various drug concentrations. (T) PGE2 levels in cell culture supernatant of BMDMs. (U) PGE2 levels in cell culture supernatant of BMDMs when using SW033291 inhibitors. Statistical analysis was performed by unpaired Student's t test (C-F, N-P, R-T) or one-way ANOVA followed by Tukey's multiple comparison test (G-H, J-M, Q, U). The data are presented as means ± SD (n = 3–6). *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001

Article Snippet: The PGE2 levels in supernatants were analyzed by the PGE2 ELISA Kit (MULTI Sciences, Cat. EK8103/2–96).

Techniques: Gene Expression, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Expressing, Cell Culture, Comparison

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NitraTh epitope-based neoantigen vaccines for effective tumor immunotherapy

doi: 10.1007/s00262-024-03830-2

Figure Lengend Snippet: The following schematic diagram illustrates the mechanism of NitraTh epitope-based neoantigen vaccines. The NitraTh epitope-based neoantigen vaccines have dual activity: 1) inducing stronger CTL and immune memory by activating CD4 + T cells, 2) deregulating immunosuppression in the tumor microenvironment by modulating macrophages to reduce PGE2 levels

Article Snippet: The PGE2 levels in supernatants were analyzed by the PGE2 ELISA Kit (MULTI Sciences, Cat. EK8103/2–96).

Techniques: Vaccines, Activity Assay

Journal: Aging Cell

Article Title: Decreased PTGES2 Farnesylation in Granulosa Cells Compromises PGE2 ‐Dependent Cumulus Expansion and Oocyte Maturation During Ovarian Aging

doi: 10.1111/acel.70374

Figure Lengend Snippet: Farnesylated PTGES2 is essential for cumulus expansion and oocyte maturation. (A) Metabolic labeling of cells with alk‐FOH reporter and subsequent CuAAC ligation with bioorthogonal detection tags for proteomics. (B) Overlap of twice‐acquired farnesylated proteomics. (C) Overlap of farnesylated proteins based on our data and the previous report (Charron et al. ). (D) Western blotting showing the expression of PTGES2 in the CTL, alk‐FOH, and alk‐FOH + FOH groups. (E) Mean gray value of PTGES2 in the CTL, alk‐FOH, and alk‐FOH + FOH groups. CTL (control) group: KGN cells treated without alk‐FOH or FOH; alk‐FOH group: KGN cells treated with 50 μM alk‐FOH; alk‐FOH + FOH group: KGN cells treated with 50 μM alk‐FOH + 50 μM FOH. (F) Western blotting shows the expression of PTGES2 in the CTL, OE, and C16S groups. (G) Mean gray value of PTGES2 in the CTL, OE, and C16S groups. CTL (control) group: 293 T cells transfected with vehicle plasmid; OE group: 293 T cells transfected with the PTGES2 overexpressing plasmid and treated with 50 μM alk‐FOH; C16S group: 293 T cells transfected with the PTGES2 C16S mutant plasmid and treated with 50 μM alk‐FOH. alk‐FOH, alkynyl‐farnesol. CuAAC, Cu‐catalyzed azide‐alkyne cycloaddition. PTGES2, prostaglandin E2 synthase 2. PD, pull down. TL, Total. His, 6 × hexahistidine tag. Data are shown as means ± SD from at least three independent repeats. Statistical analysis was performed one‐way ANOVA. ** p < 0.01, *** p < 0.001.

Article Snippet: The PGE2 levels were measured via a PGE2 ELISA Kit (Elabscience, E‐EL‐0034) according to the manufacturer's instructions.

Techniques: Labeling, Ligation, Western Blot, Expressing, Control, Transfection, Plasmid Preparation, Mutagenesis

Journal: Aging Cell

Article Title: Decreased PTGES2 Farnesylation in Granulosa Cells Compromises PGE2 ‐Dependent Cumulus Expansion and Oocyte Maturation During Ovarian Aging

doi: 10.1111/acel.70374

Figure Lengend Snippet: PTGES2 farnesylation facilitates localization to the endoplasmic reticulum and PGE2 production. (A) Western blotting showing PTGES2 expression in the membrane fractions in the CTL, FOH, and FOH + FTI‐277 groups. (B) Mean gray value of PTGES2 in the CTL, FOH, and FOH + FTI‐277 groups. (C) Immunofluorescence co‐staining of PTGES2 and calnexin in the CTL, FOH, and FOH + FTI‐277 groups. Scale bars, 10 μm. (D) Relative fluorescent intensity of PTGES2 and calnexin co‐staining in the CTL, FOH, and FOH + FTI‐277 groups. CTL (control) group: KGN cells without treatment; alk‐FOH group: KGN cells treated with 50 μM alk‐FOH; alk‐FOH + FOH group: KGN cells treated with 50 μM alk‐FOH + 50 μM FOH. (E) Western blotting showing PTGES2 expression in the membrane fractions in the CTL, OE, and C16S groups. (F) Mean gray value of PTGES2 in the CTL, OE, and C16S groups. CTL (control) group: 293 T cells transfected with vehicle plasmid; OE group: 293 T cells transfected with the PTGES2 overexpressing plasmid and treated with 50 μM alk‐FOH; C16S group: 293 T cells transfected with the PTGES2 C16S mutant plasmid and treated with 50 μM alk‐FOH. (G) ELISA showing PGE2 levels in the conditional culture media of KGN cells in the CTL, FOH, and FOH + FTI‐277 groups. FOH, farnesol. PTGES2, prostaglandin E2 synthase 2. OE, PTGES2‐overexpression plasmid. C16S, single‐point mutation plasmid of the CaaX motif in PTGES2. Data are shown as means ± SD from at least three independent repeats. Statistical analysis was performed via one‐way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The PGE2 levels were measured via a PGE2 ELISA Kit (Elabscience, E‐EL‐0034) according to the manufacturer's instructions.

Techniques: Western Blot, Expressing, Membrane, Immunofluorescence, Staining, Control, Transfection, Plasmid Preparation, Mutagenesis, Enzyme-linked Immunosorbent Assay, Over Expression

Journal: Aging Cell

Article Title: Decreased PTGES2 Farnesylation in Granulosa Cells Compromises PGE2 ‐Dependent Cumulus Expansion and Oocyte Maturation During Ovarian Aging

doi: 10.1111/acel.70374

Figure Lengend Snippet: Farnesylation of PTGES2 improves aged cumulus expansion and oocyte maturation through PGE2. (A) Representative COC images before and after cumulus expansion in the Young, Old, Old + FOH, and Old + PGE2 groups. Scale bars, 100 μm. (B) COC diameter before cumulus expansion in the Young ( n = 16), Old ( n = 16), Old + FOH ( n = 14), and Old + PGE2 ( n = 15) groups. (C) Fold change of COC diameter before and after cumulus expansion in the Young ( n = 16), Old ( n = 16), Old + FOH ( n = 14), and Old + PGE2 ( n = 15) groups. (D) Representative oocyte images in the Young, Old, Old + FOH, and Old + PGE2 groups. Scale bars, 100 μm. (E) PBE rates of oocytes in the Young ( n = 46), Old ( n = 41), Old + FOH ( n = 40), and Old + PGE2 ( n = 45) groups. (F) Representative images of spindle morphologies and chromosome alignment of oocytes in the Young, Old, Old + FOH, and Old + PGE2 groups. Scale bars, 25 μm. (G) Meiotic defect rates of oocytes in the Young ( n = 34), Old ( n = 36), Old + FOH ( n = 31), and Old + PGE2 ( n = 33) groups. (H) Representative images of 2‐cell embryos from MIIOs in the Young, Old, Old + FOH, and Old + PGE2 groups. (I) The 2‐cell embryos rate in the Young ( n = 46), Old ( n = 40), Old + FOH ( n = 38), and Old + PGE2 ( n = 36). COCs, cumulus‐oocyte complexes. FOH, farnesol. PGE2, prostaglandin E2. IVM, in vitro maturation, PBE, polar body extrusion. Young group: Young COCs cultured in MEMα; Old group: Old COCs cultured in MEMα; Old + FOH group: Old COCs cultured in MEM supplemented with 50 μM FOH; Old + PGE2 group: Old COCs cultured in MEM supplemented with 1 μM PGE2. Data are shown as means ± SD from at least three independent repeats. Statistical analysis was performed via one‐way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, *** p < 0.0001, ns, not significant.

Article Snippet: The PGE2 levels were measured via a PGE2 ELISA Kit (Elabscience, E‐EL‐0034) according to the manufacturer's instructions.

Techniques: In Vitro, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Suppression of Cytosolic Phospholipase A2 in the Ventromedial Hypothalamus Induces Hyperphagia and Obesity in Male Mice

doi: 10.3390/ijms26157532

Figure Lengend Snippet: Insulin injection increased phospho-ERK expression but not phospho-cPLA2 expression in male mice. ( a ) Representative images of phospho-ERK (p-ERK) immunohistochemistry in the hypothalamus 60 min after i.p. injecton of saline ( n = 4) or 0.75 U/kg insulin ( n = 3). ( b ) Quantification of p-ERK expression in the hypothalamus; ARC: arcuate nucleus; VMH: ventromedial hypothalamus. ( c ) Representative images of phospho-cPLA2 (p-cPLA2) immunohistochemistry in the hypothalamus 60 min after i.p. injection of saline ( n = 4) or 0.75 U/kg insulin ( n = 4). ( d ) Quantification of p-ePLA2 expression in the hypothalamus. ( e ) Prostaglandin E2 levels in the hypothalamus of shSCRM ( n = 4) or shPLA2 ( n = 5) injected mice. Arrowhead: p-ERK immunoreactivity. Scale bars: 200 µm. All data represent the mean ± SEM; **** p < 0.0001. The unpaired Student’s t -test (two-tailed p -value) was used for statistical analysis.

Article Snippet: Tissue PGE2 levels were determined by ELISA (PGE2 ELISA Kit, Elabscience, Houston, TX, USA), with absorbance measured at 450 nm using a microplate reader (SH9000Lab, Corona Electric, Hitachinaka, Japan).

Techniques: Injection, Expressing, Immunohistochemistry, Saline, Two Tailed Test