Journal: Cancers

Article Title: Aminopeptidase Expression in Multiple Myeloma Associates with Disease Progression and Sensitivity to Melflufen

doi: 10.3390/cancers13071527

Figure Lengend Snippet: List of evaluated aminopeptidase genes ( n = 39), sorted by median log2(RPKM) expression from RNAseq data from 122 multiple myeloma patient samples in the FIMM dataset.

Article Snippet: Frozen solutions containing the aminopeptidases LAP3 (TP309052) (OriGene, Rockville, MD, USA), DPP3 (8087), DPP7 (3438-SE), LTA4H (4008-ZN), RNPEP (8089-ZN), XNPEP1 (2970-ZN), and ANPEP (3815-ZN) (R&D Systems, Minneapolis, MN, USA) were thawed and diluted with their incubation buffers (Online ) to the desired concentrations.

Techniques: Expressing, Membrane, Coagulation

Journal: Cancers

Article Title: Aminopeptidase Expression in Multiple Myeloma Associates with Disease Progression and Sensitivity to Melflufen

doi: 10.3390/cancers13071527

Figure Lengend Snippet: Aminopeptidases LAP3, LTA4H, and RNPEP can hydrolyze melflufen to melphalan and 4-F-Phe-OEt. ( A ) Reaction pathway of the hydrolysis of melflufen peptide bond by the aminopeptidases LAP3, LTA4H, and RNPEP. ( B – D ) Incubation buffer and substrates were mixed and incubated without and with ( B ) LAP3, ( C ) LTA4H, and ( D ) RNPEP for 2 h at 37 °C (with duplicate incubations). The incubation was stopped by adding ice-cold acetonitrile. Using LC-HRMS, 4-F-Phe-OEt was then quantified, with concentrations presented as the peak area in the figure. 4-F-Phe-OEt: para-fluoro-L-phenylalanine ethyl ester; LC-HRMS: liquid chromatography–high-resolution mass spectrometry.

Article Snippet: Frozen solutions containing the aminopeptidases LAP3 (TP309052) (OriGene, Rockville, MD, USA), DPP3 (8087), DPP7 (3438-SE), LTA4H (4008-ZN), RNPEP (8089-ZN), XNPEP1 (2970-ZN), and ANPEP (3815-ZN) (R&D Systems, Minneapolis, MN, USA) were thawed and diluted with their incubation buffers (Online ) to the desired concentrations.

Techniques: Incubation, Liquid Chromatography, Mass Spectrometry

Journal: International Journal of Molecular Sciences

Article Title: How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females ( Cervus elaphus L.) in Different Reproductive Stages?

doi: 10.3390/ijms24054771

Figure Lengend Snippet: mRNA expression of 5-LO ( A ), LTA4H ( B ), LTC4S ( C ), PTGS2 ( D ), PGES ( E ), PGFS ( F ), and PGIS ( G ) in uterine tissues (endometrium and myometrium) on 4th and 13th day of the estrous cycle, and in pregnancy and anestrus phase. Data were normalized against GAPDH for mRNA expression using GraphPad PRISM (Version 8.3.0). Each bar represents one experimental group with SEM. Statistical differences were analyzed withy two-way analysis (ANOVA) of variance followed by Tukey’s post hoc test. The lowest statistical significance was p < 0.05. Asterisks indicate statistical differences between endometrium and myometrium (* p < 0.05; ** p < 0.01; *** p < 0.001). Different letters indicate statistical differences ( p < 0.05) between the experimental reproductive stages throughout endometrium (a, b) and myometrium (A, B), respectively.

Article Snippet: Afterward, the proteins were transferred to 0.2 μm nitrocellulose membranes in transfer buffer by electroblotting for 1.5 h. The membranes were blocked in a 5% solution of skimmed milk with 1x TBS-T for 1.5 h at room temperature (RT), and then incubated overnight at 4 °C with specific primary antibodies for 5-LO (Cayman, 160402, 1:1000), LTA4H (Cayman, 160250, 1:1000), LTC4S (Biorbyt, orb580627, 1:1000), PTGS2 (Cayman, 160107, 1:1000), PGES (Cayman, 160140, 1:1000), PGFS (Cayman, 10007940, 1:1000), and PGIS (Cayman, 160640, 1:1000).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females ( Cervus elaphus L.) in Different Reproductive Stages?

doi: 10.3390/ijms24054771

Figure Lengend Snippet: Protein expression of 5-LO ( A ), LTA4H ( B ), LTC4S ( C ), PTGS2 ( D ), PGES ( E ), PGFS ( F ), and PGIS ( G ) in uterine tissues (endometrium and myometrium) on 4th and 13th day of the estrous cycle, and in pregnancy and anestrus phase. Data were normalized against ACTB for protein expression using GraphPad PRISM (Version 8.3.0). Each bar represents one experimental group with SEM. Statistical differences were analyzed with two-way analysis (ANOVA) of variance followed by Tukey’s post hoc test. The lowest statistical significance was p < 0.05. Asterisks indicate statistical differences between endometrium and myometrium (* p < 0.1; *** p < 0.001). Different letters indicate statistical differences ( p < 0.05) between the experimental reproductive stages throughout endometrium (a, b) and myometrium (A, B), respectively.

Article Snippet: Afterward, the proteins were transferred to 0.2 μm nitrocellulose membranes in transfer buffer by electroblotting for 1.5 h. The membranes were blocked in a 5% solution of skimmed milk with 1x TBS-T for 1.5 h at room temperature (RT), and then incubated overnight at 4 °C with specific primary antibodies for 5-LO (Cayman, 160402, 1:1000), LTA4H (Cayman, 160250, 1:1000), LTC4S (Biorbyt, orb580627, 1:1000), PTGS2 (Cayman, 160107, 1:1000), PGES (Cayman, 160140, 1:1000), PGFS (Cayman, 10007940, 1:1000), and PGIS (Cayman, 160640, 1:1000).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females ( Cervus elaphus L.) in Different Reproductive Stages?

doi: 10.3390/ijms24054771

Figure Lengend Snippet: List of monoclonal antibodies used for labeling lymphocytes for flow cytometry.

Article Snippet: Afterward, the proteins were transferred to 0.2 μm nitrocellulose membranes in transfer buffer by electroblotting for 1.5 h. The membranes were blocked in a 5% solution of skimmed milk with 1x TBS-T for 1.5 h at room temperature (RT), and then incubated overnight at 4 °C with specific primary antibodies for 5-LO (Cayman, 160402, 1:1000), LTA4H (Cayman, 160250, 1:1000), LTC4S (Biorbyt, orb580627, 1:1000), PTGS2 (Cayman, 160107, 1:1000), PGES (Cayman, 160140, 1:1000), PGFS (Cayman, 10007940, 1:1000), and PGIS (Cayman, 160640, 1:1000).

Techniques: Labeling, Cytometry, Sequencing, Amplification

Journal: International Journal of Molecular Sciences

Article Title: How Is Arachidonic Acid Metabolism in the Uterus Connected with the Immune Status of Red Deer Females ( Cervus elaphus L.) in Different Reproductive Stages?

doi: 10.3390/ijms24054771

Figure Lengend Snippet: Oligonucleotide sequences used for real-time PCR. GAPDH—Glyceraldehyde 3-phosphate dehydrogenase, RN18S1—18S ribosomal RNA, ACTB—β-actin, 5-LO—5-Lipoxygenase, LTA4H—Leukotriene A4 hydrolase, LTC4S—Leukotriene C4 Synthase, PTGS2—prostaglandin endoperoxide synthase 2, PGES—Prostaglandin E2 synthase, PGFS—Prostaglandin F2alpha synthase, PGIS—Prostaglandin I2 synthase.

Article Snippet: Afterward, the proteins were transferred to 0.2 μm nitrocellulose membranes in transfer buffer by electroblotting for 1.5 h. The membranes were blocked in a 5% solution of skimmed milk with 1x TBS-T for 1.5 h at room temperature (RT), and then incubated overnight at 4 °C with specific primary antibodies for 5-LO (Cayman, 160402, 1:1000), LTA4H (Cayman, 160250, 1:1000), LTC4S (Biorbyt, orb580627, 1:1000), PTGS2 (Cayman, 160107, 1:1000), PGES (Cayman, 160140, 1:1000), PGFS (Cayman, 10007940, 1:1000), and PGIS (Cayman, 160640, 1:1000).

Techniques: Real-time Polymerase Chain Reaction