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Elabscience Biotechnology pge2 levels

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, <t>prostaglandin</t> <t>E2</t> 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.

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1) Product Images from "Decreased PTGES2 Farnesylation in Granulosa Cells Compromises PGE2 ‐Dependent Cumulus Expansion and Oocyte Maturation During Ovarian Aging"

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

Journal: Aging Cell

doi: 10.1111/acel.70374

Figure Legend 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.

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

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

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

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.

Figure Legend 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.

Techniques Used: In Vitro, Cell Culture

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(A) Immunostaining of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h. (B) Immunoblot analysis of inactive and active forms of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h. (C) Enzymatic activity of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL. * P =0.025 for ATCC 17978 vs. CTL, ** P =0.05 for ATCC 17978 vs. ATCC 1797 8 + BEL (5 µM), *** P =0.042 for ATCC 17978 vs. ATCC 17978 + BEL (10 µM). (D) Cox-2 enzymatic activity in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL and NS. (* P =0.0011 for ATCC 17978 vs. CTL, * P =0.049 for CTL vs. ATCC 17978 + BEL (5 µM), * P =0.0096 for CTL vs. ATCC 17978 + NS, ** P =0.0005 for ATCC 17978 vs. ATCC 17978 + BEL (5 µM), *** P =0.001 for ATCC 17978 vs. ATCC 17978 + NS. ( E ) PGE2 production in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL and NS. * P =0.004 for ATCC 17978 vs. CTL, ** P =0.0417 for ATCC 17978 vs. ATCC 17978 + BEL (5 µM), *** P =0.0015 for ATCC 17978 vs. ATCC 17978 + NS. CTL: control; Str: staurosporine; BEL: bromoelone lactone; NS: cyclooxygenase inhibitor; Cox-2: cyclooxygenase-2; PGE2: prostaglandin 2.

Journal: bioRxiv

Article Title: OmpA controls intracellular survival of Acinetobacter baumannii through TFEB activation and lysosomal remodeling

doi: 10.64898/2026.04.18.719357

Figure Lengend Snippet: (A) Immunostaining of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h. (B) Immunoblot analysis of inactive and active forms of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h. (C) Enzymatic activity of iPLA 2 in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL. * P =0.025 for ATCC 17978 vs. CTL, ** P =0.05 for ATCC 17978 vs. ATCC 1797 8 + BEL (5 µM), *** P =0.042 for ATCC 17978 vs. ATCC 17978 + BEL (10 µM). (D) Cox-2 enzymatic activity in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL and NS. (* P =0.0011 for ATCC 17978 vs. CTL, * P =0.049 for CTL vs. ATCC 17978 + BEL (5 µM), * P =0.0096 for CTL vs. ATCC 17978 + NS, ** P =0.0005 for ATCC 17978 vs. ATCC 17978 + BEL (5 µM), *** P =0.001 for ATCC 17978 vs. ATCC 17978 + NS. ( E ) PGE2 production in HeLa cells infected with the ATCC 17978 strain for 24 h, in the presence or absence of BEL and NS. * P =0.004 for ATCC 17978 vs. CTL, ** P =0.0417 for ATCC 17978 vs. ATCC 17978 + BEL (5 µM), *** P =0.0015 for ATCC 17978 vs. ATCC 17978 + NS. CTL: control; Str: staurosporine; BEL: bromoelone lactone; NS: cyclooxygenase inhibitor; Cox-2: cyclooxygenase-2; PGE2: prostaglandin 2.

Article Snippet: Prostaglandin E2 (PGE2) levels were quantified in three independent experiments using the PGE2-EIA monoclonal kit (Cayman Chemical, USA) from culture supernatants collected after the same treatments.

Techniques: Immunostaining, Infection, Western Blot, Activity Assay, Control

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

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.

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

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