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: Inhibition of farnesylation leads to cumulus expansion and oocyte maturation impairment in young COCs in vitro. (A) A schematic diagram showing young COCs collection and FTI‐277 treatment for analyzing cumulus expansion and oocyte maturation. (B) Representative COC images before and after cumulus expansion in the CTL and FTI‐277 groups. Scale bars, 100 μm. (C) COC diameter analysis before cumulus expansion in the CTL ( n = 19) and FTI‐277 ( n = 25) groups. (D) Fold change of COC diameter before and after cumulus expansion in the CTL ( n = 19) and FTI‐277 ( n = 25) groups. (E) Representative oocyte images in the CTL and FTI‐277 groups. Scale bars, 100 μm. (F) PBE rates of oocytes in the CTL ( n = 36) and FTI‐277 ( n = 39) groups. (G) Representative images of spindle morphologies and chromosome alignment of oocytes in the CTL and FTI‐277 groups. Scale bars, 25 μm. (H) Meiotic defect rates of oocytes in the CTL ( n = 33) and FTI‐277 ( n = 34) groups. COCs, cumulus‐oocyte complexes. IVM, in vitro maturation. PBE, polar body extrusion. CTL (control) group: Young COCs cultured in MEMα. FTI‐277 group: Young COCs cultured in MEM supplemented with 50 μM FTI‐277. Data are shown as means ± SD from at least three independent repeats. Statistical analysis was performed via an unpaired Student's t‐test. ** p < 0.01, **** p < 0.0001, ns, not significant.

Article Snippet: The concentration of FOH, FTI‐277 (Aladdin, F331609), and PGE2 (MCE, HY‐101952) used in this study was 50, 50, and 1 μM, respectively.

Techniques: Inhibition, In Vitro, Control, Cell Culture

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: FOH supplementation improves cumulus expansion and oocyte maturation in aged COCs via farnesylation in vitro. (A) A schematic diagram showing old COCs collection and treatment with FOH or FOH + FTI‐277 for analyzing cumulus expansion and oocyte maturation. (B) Representative COC images before and after cumulus expansion in the CTL, FOH, and FOH + FTI‐277 groups. Scale bars, 100 μm. (C) COC diameter analysis before cumulus expansion in the CTL ( n = 14), FOH ( n = 14), and FOH + FTI‐277 ( n = 12) groups. (D) Fold change of COC diameter before and after cumulus expansion in the CTL ( n = 14), FOH ( n = 14), and FOH + FTI‐277 ( n = 12) groups. (E) Representative oocyte images in the CTL, FOH, and FOH + FTI‐277 groups. Scale bars, 100 μm. (F) PBE rates of oocytes in the CTL ( n = 36), FOH ( n = 42), and FOH + FTI‐277 ( n = 40) groups. (G) Representative images of spindle morphologies and chromosome alignment of oocytes in the CTL, FOH, and FOH + FTI‐277 groups. Scale bars, 25 μm. (H) Meiotic defect rates of oocytes in the CTL ( n = 30), FOH ( n = 37), and FOH + FTI‐277 ( n = 44) groups. COCs, cumulus‐oocyte complexes. FOH, farnesol. IVM, in vitro maturation. PBE, polar body extrusion. CTL (control) group: Old COCs cultured in MEMα; FOH group: Old COCs cultured in MEM supplemented with 50 μM FOH; FOH + FTI‐277 group: Old COCs cultured in MEMα supplemented with 50 μM FOH + 25 μM FTI‐277. Data are shown as means ± SD from at least three independent repeats. Statistical analysis was performed via one‐way ANOVA. ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant.

Article Snippet: The concentration of FOH, FTI‐277 (Aladdin, F331609), and PGE2 (MCE, HY‐101952) used in this study was 50, 50, and 1 μM, respectively.

Techniques: In Vitro, Control, Cell Culture

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 concentration of FOH, FTI‐277 (Aladdin, F331609), and PGE2 (MCE, HY‐101952) used in this study was 50, 50, and 1 μM, respectively.

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

Journal: Chemical Biology & Drug Design

Article Title: Investigation on the antipyretic mechanism of Chaiqin Qingning capsule for the treatment of fever based on network pharmacology, molecular docking, and in vitro experimental validation

doi: 10.1111/cbdd.14451

Figure Lengend Snippet: FIGURE 7 Western blot detection of CQQNC on the expression of COX-1, COX-2, cAMP, EP3, and mPGES1. Statistical significance: ∆∆∆∆p < .0001, ∆∆p < .01, and ∆p < .05 represents IL-1β group versus control group. ****p < .0001, **p < .01, and *p < .05 represent CQQNC group versus IL-1β group (n = 3).

Article Snippet: AntiCOX- 1 (batch number: 00100314), COX- 2 (batch number: 10027896), EP3 (batch number: 00071265), mouse and rabbit GAPDH (batch number: 10021642 and 0013796) antibody were purchased from Proteintech.

Techniques: Western Blot, Expressing, Control

Journal: Cell communication and signaling : CCS

Article Title: PTGER4 signaling regulates class IIa HDAC function and SPINK4 mRNA levels in rectal epithelial cells.

doi: 10.1186/s12964-024-01879-1

Figure Lengend Snippet: Fig. 5 PGE2-PTGER4 activates HDACs to increase SPINK4 expression. Western blot analysis for A EP4i, B PGE2 or EP4i, or C butyrate-treated organoid protein lysates probed for HDAC4 (S246)/ HDAC5 (S259)/ HDAC7 (S155), HDAC 4, and GAPDH as a loading control. The experiment in panel A is representative of an experiment that was performed on 2 different patients. Panels B and C are representative of experiments performed on 3 different patient samples. D RT-PCR fold-change quantification of SPINK4 normalized to β-ACTIN in organoids treated with PGE2 and/or LMK-235 (n = 5 different patient samples were used in this experiment). ns = non-significant; *P < 0.05; **P < 0.01; Two-way ANOVA

Article Snippet: For detecting PTGER4 by Western blotting, rabbit anti-PTGER4 was obtained from Proteintech (cat. no. 24895–1-AP).

Techniques: Expressing, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction

Journal: Animals : an Open Access Journal from MDPI

Article Title: FABP3 Mediates Lipid Droplet Accumulation and Adhesive Capacity in Bovine Endometrial Epithelial Cells via PGE 2 /PTGER4/PPAR Axis

doi: 10.3390/ani15233417

Figure Lengend Snippet: PGE 2 binds to PTGER4 to regulate FABP3 expression in bEECs: ( A ) The difference in PTGER ( PTGER1 , PTGER2 , PTGER3 and PTGER4 ) expression in bEECs after treatment with P 4 and/or PGE 2 . ( B – D ) The representative images and analysis of PTGER2 and PTGER4 expression in bEECs after treatment with P 4 and/or PGE 2 . ( E , F ) The representative images and analysis of PTGER2 expression in bEECs between shN and shPTGER2 groups after treatment with DMSO or P 4 + PGE 2 . ( G , H ) The representative images and analysis of PTGER4 expression in bEECs between shN and shPTGER4 groups after treatment with DMSO or P 4 + PGE 2 . ( I ) The analysis of FABP3 expression in bEECs after treatment with DMSO, PGE 2 or the inhibitor of PTGER4 ( GW627368 ). The data are means ± SEM of three independent experiments. Statistically significant differences are indicated by asterisks: ** p < 0.01, ns p > 0.05. Bars with different letters are significantly different ( p < 0.05).

Article Snippet: Then, PGE 2 (10 −8 –10 −6 M; 14010, Cayman, Ann Arbor, MI, USA) was added to the medium, and the cells were cultured for 24 h. For inhibitor treatment, 5 and 10 μM PTGER2 inhibitor (TG-155; MedChemExpress, Newark, NJ, USA) or 5 and 10 μM PTGER4 inhibitor ( GW627368 ; Selleck, Houston, TX, USA) were added to the medium 1 h prior to the PGE 2 treatment.

Techniques: Expressing