Journal: bioRxiv

Article Title: Functional characterization of eicosanoid signaling in Drosophila development

doi: 10.1101/2025.01.13.632770

Figure Lengend Snippet: (A) Unrooted maximum-likelihood phylogenetic tree of class A G protein-coupled receptors (GPCRs) in H. sapiens and D. melanogaster . GPCRs in D. melanogaster are indicated by blue branches, whereas branches for H. sapiens GPCRs are color-coded based on different classes of ligands. The scale bar indicates an evolutionary distance of 0.5 amino acid substitutions per site. Opsins and leucine-rich repeat containing GPCRs are not included in the phylogenetic tree. Accession numbers of the receptors analyzed are listed in S1 Table. (B) List of lipid mediator receptors in H. sapiens and their Drosophila orthologs. (C) Luminescence responses of S2 cells co-expressing Aequorin and G α 15 , along with CG7497/PGR or H. sapiens PGE2 receptor EP2 ( HsEP2 ), to prostanoids or their synthetic analogs. Relative luminescence normalized by luminescence in ligand-free controls is shown. Cells expressing CG7497/PGR were significantly activated by all ligands, whereas cells expressing EP2 were activated only by PGE2. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 (Dunnett’s test vs mock treatment control).

Article Snippet: Early pupae (up to 24 h APF) were collected and kept in Drosophila narrow vials (Genesee Scientific) containing wet tissue at 25°C under 12 h-light and 12 h-dark photoperiod.

Techniques: Expressing, Control

Journal: bioRxiv

Article Title: Functional characterization of eicosanoid signaling in Drosophila development

doi: 10.1101/2025.01.13.632770

Figure Lengend Snippet: (A) Schematic diagram of prostanoid synthetic pathways. Multiple prostanoids are synthesized from arachidonic acid (AA) by different enzymes. (B) Drosophila enzymes orthologous to human prostanoid synthases were identified by phylogenetic analyses. Phylogenetic trees are shown in S4–S10 Figs. PGD, PGE, and PGF synthases are highly conserved in insects, although D. melanogaster lacks some of them. (C) Prostanoids detected in whole body extracts of pupae by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based lipidomics. AA was detected in intact pupae at 0–4 hours after pupation, whereas prostanoids were undetectable. PGD2, PGE2, and PGF2α were detected in pupae that were raised in AA-supplemented diet during larval development. Data shown are mean total amount of eicosanoids in the whole body per animal ± SEM. n = 4–5.

Article Snippet: Early pupae (up to 24 h APF) were collected and kept in Drosophila narrow vials (Genesee Scientific) containing wet tissue at 25°C under 12 h-light and 12 h-dark photoperiod.

Techniques: Synthesized, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy

Journal: bioRxiv

Article Title: Functional characterization of eicosanoid signaling in Drosophila development

doi: 10.1101/2025.01.13.632770

Figure Lengend Snippet: (A) Developmental phenotype of PGD and PGE synthase ortholog mutants. Null mutant of cPges / p23 and triple mutants of Mgst - , CG33177 , and CG33178 died during pupa-adult development. Multiple comparison Chi-square test with Bonferroni correction was applied among cPges / p23 mutants including the heterozygous control. *** p < 0.001. (B) Representative images of cPges / p23 knockout flies. About 70% of flies died before wing pigmentation, whereas ∼30% of flies died as pharate adults. Scale bar, 200 µm. (C, D) Relative expression levels of hypoxia response genes, Ldh (C) and Hph (D), in the PGD and PGE synthase ortholog mutants. The cPges / p23 mutant showed high expression of hypoxia response genes at 24 and 48 hours after puparium formation (APF). Expression levels are normalized by the levels of a reference gene, rp49 , in the same cDNA samples and shown as relative to 0 hours APF of GstS1 - . n = 3–4. p < 0.001 (Tukey’s test). (E) Rescue of the cPges / p23 mutant by Gal4 -driven expression of either cPges / p23 or human prostaglandin E synthase 3 ( HsPTGES3 ). The lethal phenotype was rescued by overexpressing cPges / p23 or HsPTGES3 either ubiquitously ( tubP-Gal4 ), in PGR expression sites ( PGR-Gal4 ), in tracheal cells ( btl-Gal4 ), or in the epidermis ( Eip71CD-Gal4 ). ** p < 0.01, *** p < 0.001 (Chi-square test vs + > cPges / p23 or + > HsPTGES3 with Bonferroni correction). (F) Developmental deficiency caused by cPges / p23 RNAi. cPges / p23 knockdown flies died during pupa-adult development when induced using either a ubiquitous driver (tubP-Gal4 ) or a combination of the tracheal cell ( btl-Gal4 ) and epidermis ( Eip71CD-Gal4 ) drivers. Lethality caused by cPges / p23 RNAi in tracheal cells and epidermis was significantly rescued by 40% O 2 . *** p < 0.001 (multiple comparison Chi-square test vs + > cPges / p23 RNAi with Bonferroni correction, or 20% O 2 vs 40% O 2 ). (G) PGE2 conversion assay using Drosophila S2 cells. Arachidonic acid (AA) was added to the culture medium of S2 cells transfected with cPges / p23 or HsPTGES3 along with a H. sapiens cyclooxygenase ( HsCOX1 or HsCOX2 ), and PGE2 amount in the medium was determined. The basal levels of PGE2 immunoactivity in all samples is likely due to the cross-reactivity of the antibody used in the assay. Increased PGE2 immunoreactivity was detected in cyclooxygenase -expressing cells regardless of cPges / p23 or HsPTGES3 transfection. n = 4. (H) Reduction of the PGE2 synthetic activity of S2 cells by knocking down endogenous cPges / p23 expression. AA was added to the culture medium of S2 cells transfected with HsCOX2 and treated with cPges / p23 dsRNAs. PGE2 amount in the medium was determined. n = 2 (AA 0 µM) or n = 4 (AA 10 µM). *** p < 0.001 (Dunnett’s test vs EGFP dsRNA-treated cells).

Article Snippet: Early pupae (up to 24 h APF) were collected and kept in Drosophila narrow vials (Genesee Scientific) containing wet tissue at 25°C under 12 h-light and 12 h-dark photoperiod.

Techniques: Mutagenesis, Comparison, Control, Knock-Out, Expressing, Knockdown, Transfection, Activity Assay

Journal: STAR Protocols

Article Title: A simplified courtship conditioning protocol to test learning and memory in Drosophila

doi: 10.1016/j.xpro.2022.101572

Figure Lengend Snippet:

Article Snippet: Drosophila Anti-Mite Vial Plugs (Droso-Plugs®)∗∗ , Genesee Scientific , 59–200, 59–201, 59–194.

Techniques: Recombinant, Software, Cell Culture

Journal: Molecules

Article Title: Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study

doi: 10.3390/molecules28062752

Figure Lengend Snippet: Positive grading quantification of immunocytochemical staining of SOD1 in the MCF-7/WT line under the influence of compounds. The number of stained cells was determined by counting 100 cells in three randomly selected fields. The intensity of the immunohistochemical staining was assessed as (-) negative (no reaction), (+) weak, (++) moderate, and (+++) strong.

Article Snippet: A primary antibody was used: anti-SOD1 rabbit polyclonal antibody (orb39428, Biorbyt, Cambridge, UK).

Techniques: Staining, Immunohistochemical staining, Control

Journal: Molecules

Article Title: Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study

doi: 10.3390/molecules28062752

Figure Lengend Snippet: Positive grading quantification of immunocytochemical staining of SOD1 in the MCF-7/WT line under the influence of 17β-E2 with Cr(VI). The number of stained cells was determined by counting 100 cells in three randomly selected fields. The intensity of the immunohistochemical staining was assessed as (-) negative (no reaction), (+) weak, (++) moderate, and (+++) strong.

Article Snippet: A primary antibody was used: anti-SOD1 rabbit polyclonal antibody (orb39428, Biorbyt, Cambridge, UK).

Techniques: Staining, Immunohistochemical staining, Control

Journal: Molecules

Article Title: Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study

doi: 10.3390/molecules28062752

Figure Lengend Snippet: Positive grading quantification of immunocytochemical staining of SOD1 in the MCF-7/WT line under the influence of 2-MeOE2 with Cr(VI). The number of stained cells was determined by counting 100 cells in three randomly selected fields. The intensity of the immunohistochemical staining was assessed as (-) negative (no reaction), (+) weak, (++) moderate, and (+++) strong.

Article Snippet: A primary antibody was used: anti-SOD1 rabbit polyclonal antibody (orb39428, Biorbyt, Cambridge, UK).

Techniques: Staining, Immunohistochemical staining, Control

Journal: Molecules

Article Title: Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study

doi: 10.3390/molecules28062752

Figure Lengend Snippet: Positive grading quantification of immunocytochemical staining of SOD1 in the MCF-7/WT line under the influence of 4-OHE2 with Cr(VI). The number of stained cells was determined by counting 100 cells in three randomly selected fields. The intensity of the immunohistochemical staining was assessed as (-) negative (no reaction), (+) weak, (++) moderate, and (+++) strong.

Article Snippet: A primary antibody was used: anti-SOD1 rabbit polyclonal antibody (orb39428, Biorbyt, Cambridge, UK).

Techniques: Staining, Immunohistochemical staining, Control

Journal: Molecules

Article Title: Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study

doi: 10.3390/molecules28062752

Figure Lengend Snippet: Positive grading quantification of immunocytochemical staining of SOD1 in the MCF-7/WT line under the influence of 16α-OHE1 with Cr(VI). The number of stained cells was determined by counting 100 cells in three randomly selected fields. The intensity of the immunohistochemical staining was assessed as (-) negative (no reaction), (+) weak, (++) moderate, and (+++) strong.

Article Snippet: A primary antibody was used: anti-SOD1 rabbit polyclonal antibody (orb39428, Biorbyt, Cambridge, UK).

Techniques: Staining, Immunohistochemical staining, Control

Journal: Molecules

Article Title: Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study

doi: 10.3390/molecules28062752

Figure Lengend Snippet: Immunoassayed reaction: exemplary expression results of SOD1 in MCF-7/WT exposed to the single compounds and after exposure to the combined effects of estrogens and Cr(VI).

Article Snippet: A primary antibody was used: anti-SOD1 rabbit polyclonal antibody (orb39428, Biorbyt, Cambridge, UK).

Techniques: Expressing