Journal: Cancer Research Communications

Article Title: Inhibition of Microsomal Prostaglandin E2 Synthase Reduces Collagen Deposition in Melanoma Tumors and May Improve Immunotherapy Efficacy by Reducing T-cell Exhaustion

doi: 10.1158/2767-9764.CRC-23-0210

Figure Lengend Snippet: The influence of ptgs1 , ptgs2 , and ptges KO on the production of arachidonic acid metabolites from murine Braf V600E melanoma cells. A, Schematic diagram of the arachidonic acid metabolic cascade. PGG2, prostaglandin G2; PGH2, prostaglandin H2; TxAS, thromboxane-A synthase; TxA2, thromboxane A2; PGDS, prostaglandin D synthase; cPGES, cytosolic prostaglandin E synthase; PGFS, prostaglandin F synthase; PGIS, prostaglandin I synthase. B and C, ptgs1 , ptgs2 , and ptges genes in murine Braf V600E melanoma cells were knocked out via CRISPR/CAS9. The mRNA and protein levels were analyzed by qRT-PCR (B) and Western blotting (C). Bar graphs show the fold change relative to mRNA levels of scramble control for each gene (B, n = 4). GAPDH was used as a loading control (C). D–H, Bar plot showing the concentrations of arachidonic acid metabolites released in supernatants obtained from scramble control, ptgs1- KO, ptgs2- KO, and ptges- KO cells. All prostanoids were measured by ELISA ( n = 4): PGE2 (D), TxB2 (E), PGD2 (F), PGF2α (G), and 6-keto PGF1α (H). Graph values represent mean ± SD. Significance in difference between two groups was determined by Student t test. **, P < 0.01; *, P < 0.05.

Article Snippet: Levels of released prostaglandins and thromboxane in supernatants and tumor tissues were quantified using Prostaglandin E2 ELISA kit, Prostaglandin D2 ELISA kit, Prostaglandin F2α ELISA kit, 6-keto Prostaglandin F1α ELISA kit, and Thromboxane B2 ELISA kit (Cayman Chemical), according to the manufacturer's recommendations.

Techniques: CRISPR, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Chinese Medicine

Article Title: Rosmarinic acid treatment protects against lethal H1N1 virus-mediated inflammation and lung injury by promoting activation of the h-PGDS-PGD 2 -HO-1 signal axis

doi: 10.1186/s13020-023-00847-0

Figure Lengend Snippet: The increased expression of h-PGDS/PGD2/HO-1 was responsible for the inhibitory effects of RosA on H1N1 virus-induced NF-κB and P38 MAPK activation. A The expression of h-PGDS in H1N1 virus-infected cells was analyzed by Western blotting. B Relative protein expression of h-PGDS was normalized to GAPDH levels. C ELISA assay was performed to measure the levels of PGD 2 in the culture supernatant. D The levels of PGD 2 in the culture supernatant of h-PGDS overexpression (h-PGDS OE) plasmid-transfected A549 cells with or without H1N1 virus infection were quantified by ELISA assay. E The levels of P-p65 and P-p38 in h-PGDS overexpression (h-PGDS OE) plasmid-transfected A549 cells with or without H1N1 virus infection were detected by Western blotting. F Relative protein expression of h-PGDS, P-IKBα, P-p65 and P-p38 was normalized to GAPDH levels. G Luminex assay was performed to measure the levels of pro-inflammatory cytokines (IL-6 and TNF-α) in the culture supernatant of h-PGDS overexpression (h-PGDS OE) plasmid-transfected A549 cells with or without H1N1 virus infection. H Western blot analysis of P-IKBα, P-p65 and P-p38 in H1N1 virus-infected cells treated with RosA alone or in combination with h-PGDS inhibitor. I The relative expression of P-IKBα, P-p65 and P-p38 expression was quantified relative to GAPDH. J Luminex assay were performed to measure the levels of pro-inflammatory cytokines (IL-6, IL-8, IP-10, TNF-α, MCP-1 and RANTES) in H1N1 virus-infected cells treated with RosA alone or in combination with h-PGDS inhibitor. K The expression of HO-1 in H1N1 virus-infected cells was detected by Western blotting. L HO-1 protein levels were quantified by normalizing to GAPDH levels. M Western blot analysis of HO-1 in H1N1 virus-infected cells treated with RosA alone or in combination with h-PGDS inhibitor. N Relative HO-1 expression was quantified relative to GAPDH. O The levels of HO-1 in h-PGDS overexpression (h-PGDS OE) plasmid-transfected A549 cells with or without H1N1 virus infection were detected by Western blotting. P Relative HO-1 expression was quantified relative to GAPDH. Q Representative immunofluorescence images of h-PGDS (pink) and HO-1 (red) in lung SpC + (green) alveolar epithelial cells. R Quantitative analysis of fluorescence intensities for h-PGDS and HO-1 in SpC + alveolar epithelial cells. S The levels of PGD 2 in the lung homogenates were determined by ELISA assay. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: PGD 2 levels in the supernatant were determined by an ELISA kit (E-EL-0066, Elabscience, Wuhan, China).

Techniques: Expressing, Virus, Activation Assay, Infection, Western Blot, Enzyme-linked Immunosorbent Assay, Over Expression, Plasmid Preparation, Transfection, Luminex, Immunofluorescence, Fluorescence

Journal: bioRxiv

Article Title: Targeted Eicosanoid Profiling Reveals MicroRNA-155 Shifts PGE2/PGD2 Balance Towards Oncogenic State

doi: 10.1101/2020.04.08.032136

Figure Lengend Snippet: A. PGE2/PGD2 ratio in miR-155-positive (Lav670 and Lav714) and miR-155-depleted (Lav670+miRZIP155 and Lav714+miRZIP155) tumors (n=20). B-C. PGE2(B) and PGD2(C) level in miR-155 KO/KO and in miR-155 KO/+ tumors measured by ELISA (n=10). D-G. Relative RNA expression of Ptges1 (D), Ptges2 (E), Ptgds (F) and 15-Pgdh (G) in miR-155 KO/KO and miR-155 KO/+ tumors (n=10). H. Western blot analysis of key metabolic enzymes of PGE2/PGD2 production in miR-155 KO/KO and miR-155 KO/+ tumors (n=10). COX1, COX2, PTGES1, PTGES2, PTGDS, and 15-PGDH expressions are shown. β-actin antibody was used to ensure equal loading.

Article Snippet: The plasma concentrations of PGE2 and PGD2 were determined using ELISA kits from ENZO Life Sciences (Farmingdale, New York, NY, USA) and CUSABIO Biotechnology (Wu Han, China), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, RNA Expression, Western Blot

Journal: bioRxiv

Article Title: Targeted Eicosanoid Profiling Reveals MicroRNA-155 Shifts PGE2/PGD2 Balance Towards Oncogenic State

doi: 10.1101/2020.04.08.032136

Figure Lengend Snippet: A. Relative PGE2 (left) and PGD2 (right) levels in MDA-MB-436 and Hs-578T cells in control (in blue) or in miR-155-knockdown cells (miRZIP155 in red). B. Relative PGE2 (left) or PGD2 (right) level in MCF7 cells with overexpression of miR-155 (miRH, in yellow). C-F. Relative RNA expression of Ptges1 (C), Ptges2 (D), Ptgds(E) and 15-Pgdh (F) in MDA-MB-436, Hs-578T, and MCF7 cells after knockdown (miRZIP155, in red) or overexpression (miRH155, in yellow) of miR-155. G. Western blot analysis of key metabolic enzymes for PGE2/PGD2 production in three breast cancer cell lines with after knockdown (miRZIP155) or overexpression (miRH155) of miR-155. β-actin antibody was used to ensure equal loading

Article Snippet: The plasma concentrations of PGE2 and PGD2 were determined using ELISA kits from ENZO Life Sciences (Farmingdale, New York, NY, USA) and CUSABIO Biotechnology (Wu Han, China), respectively.

Techniques: Control, Knockdown, Over Expression, RNA Expression, Western Blot

Journal: bioRxiv

Article Title: Targeted Eicosanoid Profiling Reveals MicroRNA-155 Shifts PGE2/PGD2 Balance Towards Oncogenic State

doi: 10.1101/2020.04.08.032136

Figure Lengend Snippet: A. Relative level of miR-155 in six PDCs. Three high-expressers were labeled as PDCH and low-expressers were labeled as PDCL. Yellow bars on PDCL (miRH 155) indicates miR-155 level expressed by lentiviral infection. B and C. The level of PGE2(B) or PGD2(C) in three PDCLs with overexpressed miR-155 (miRH155, in yellow). D-G. Relative RNA expression of Ptges1 (D), Ptges2 (E), Ptgds (F) and 15-Pgdh (G) in PDCH, PDCL, and PDCL with lentiviral miR-155 expression (miRH155). H. Western blot analysis of key metabolic enzymes for PGE2/PGD2 production in three PDCH, PDCL, and PDCL with miR-155 expression (marked as +). β-actin antibody was used to ensure equal loading.

Article Snippet: The plasma concentrations of PGE2 and PGD2 were determined using ELISA kits from ENZO Life Sciences (Farmingdale, New York, NY, USA) and CUSABIO Biotechnology (Wu Han, China), respectively.

Techniques: Labeling, Infection, RNA Expression, Expressing, Western Blot

Journal: bioRxiv

Article Title: Targeted Eicosanoid Profiling Reveals MicroRNA-155 Shifts PGE2/PGD2 Balance Towards Oncogenic State

doi: 10.1101/2020.04.08.032136

Figure Lengend Snippet: A. Correlation analysis between miR-155 (in blue) and PGE2/PGD2 ratio (in red) in human TNBC specimens. p=0.045, B-E. Relative RNA expression of PTGES1 (b), PTGES2 (c), PTGDS (d) and 15-PGDH (e) in miR-155-high (H1-H10, in red) and miR-155-low (L1-L10, in blue) TNBC samples. F. Western blot results of key metabolic enzymes and suggested novel regulators for PGE2/PGD2 metabolism in the miR-155-high and -low TNBC specimens. Results of COX1, COX2, PTGES1, PTGES2, PTGDS, 15-PGDH, cMYC, and KLF4 are shown. β-actin was used to ensure equal loading. G and H. Correlation analysis between miR-155 and PGE2(g) or PGD2(h) in the plasma samples of selected TNBC patients. I. Overall survival curves of breast cancer patients according to the levels of cMYC(upper left), KLF4(upper right), PTGES2(lower left) and PTGDS(lower right), using the ProgeneV2 webtool. J. Graphical abstract showing the molecular mechanism of PGD2/PGE2 production regulated by miR-155.

Article Snippet: The plasma concentrations of PGE2 and PGD2 were determined using ELISA kits from ENZO Life Sciences (Farmingdale, New York, NY, USA) and CUSABIO Biotechnology (Wu Han, China), respectively.

Techniques: RNA Expression, Western Blot, Clinical Proteomics

Journal: iScience

Article Title: Senolysis induced by 25-hydroxycholesterol targets CRYAB in multiple cell types

doi: 10.1016/j.isci.2022.103848

Figure Lengend Snippet: 25HC induces senolysis in vivo in Doxo-treated mice (A) Timeline of 25HC treatment in Doxo-treated mice. Mice were given 4 mg/kg Doxo or PBS for 5 daily injections[A1], then injected with 25HC or vehicle for 7 consecutive days. (B) Mass of gastrocnemius muscles. Muscle weight was normalized to body weight. (C) Cdkn2a and Cryab mRNA levels in gastrocnemius (Gastroc) and soleus (Sol) muscles. RNA was collected 28–30 d after the first treatment with Doxo. mRNA levels were quantified by qRT-PCR, and normalized to actin mRNA levels. The average value of PBS-vehicle was set at 1. (D) 15-d-PGJ2 analysis in urine. Urine was collected at 24 h after the first treatment with 25HC or HbCD. p values versus PBS-Veh or Doxo-Veh are obtained using Tukey's multiple comparisons test. ∗p < 0.05, ∗∗p < 0.01, p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: 15-deoxy-D 12,14 -PGJ2 ELISA kit , Enzo Life Sciences , ADI-900-023.

Techniques: In Vivo, Injection, Quantitative RT-PCR

Journal: iScience

Article Title: Senolysis induced by 25-hydroxycholesterol targets CRYAB in multiple cell types

doi: 10.1016/j.isci.2022.103848

Figure Lengend Snippet: 25HC is an in vivo senolytic in tissues of aged mice (A) Timeline of 25HC treatment in aged mice. Young (5 months old) and old (24–26 months old) mice were treated with 25HC or vehicle for 5 consecutive days. (B) Mass of gastrocnemius muscles in male and female mice. Muscle weight was normalized to body weight. (C) Cdkn1a and Cryab mRNA levels in gastrocnemius (Gastroc) and soleus (Sol) muscles. RNA was collected at 7–9 d after the first treatment with 25HC or vehicle. mRNA levels of the indicated genes were quantified by qRT-PCR, and normalized to those of actin mRNA. The average value of young-vehicle was set at 1. (D) 15-d-PGJ2 analysis in urine. Urine was collected 12 h after the first treatment with 25HC or vehicle. p values were obtained using Tukey's multiple comparisons test. ∗p< 0.05, ∗∗p< 0.01, ∗∗∗p< 0.001∗∗∗∗p< 0.0001.

Article Snippet: 15-deoxy-D 12,14 -PGJ2 ELISA kit , Enzo Life Sciences , ADI-900-023.

Techniques: In Vivo, Quantitative RT-PCR

Journal: iScience

Article Title: Senolysis induced by 25-hydroxycholesterol targets CRYAB in multiple cell types

doi: 10.1016/j.isci.2022.103848

Figure Lengend Snippet:

Article Snippet: 15-deoxy-D 12,14 -PGJ2 ELISA kit , Enzo Life Sciences , ADI-900-023.

Techniques: Enzyme-linked Immunosorbent Assay, Sequencing