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<t>RORα</t> mediates n-3 PUFA-regulated rhythmic oscillations in molecular biology (A and B) Optimal conformation of the <t>RORα</t> <t>(PDB</t> ID: 4s15 ) docking complex bound with DHA (A) and EPA (B). The RORα structure is shown as ribbon in blue while in light blue for the enlarged zoom of the active pocket. Detailed binding residues are shown as yellow sticks, and hydrophobic interactions are all shown as semitransparent gray spheres. The yellow dotted line is hydrogen bonding, measuring a distance range of 2.0–3.0 Å; a gray translucent surface indicates hydrophobic action. (C) Free energy of binding of 4s15 receptor to ligands (4DB, DHA-derivate MaR1, DHA, and EPA) was estimated with the highest score. Molecular docking estimates were performed by AutodockVina. (D–G) Effects of 100 μM DHA/EPA plus 300 μM PA on the oscillations of circadian clock genes including Clock (D), Bmal1 (E), Per2 (F), and Rev-erb (G), in RORα-KO hypothalamic neuron cells. The relative mRNA expressions were detected at ZT 0, 4, 8, 12, 16, 20, 24, 28, and 32 after synchronization, and oscillation curves were fitted by Cosinor algorithm ( n = 3 for each group, biological replicates). (H–K) Representative immunoblotting of clock proteins including (H and J) CLOCK and (I and K) REV-ERB at ZT 4, 12, and 20 after synchronization in RORα-KO hypothalamic neuron cells ( n = 3 for each group, biological replicates). For (D)–(K), data were expressed as mean ± SEM. Statistical differences between PA treatment versus other groups at same time point were determined by one-way ANOVA followed by the Dunnett’s multiple comparisons test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Statistical differences between the same group at different time points; statistical differences were determined by one-way ANOVA followed by the Tukey’s multiple comparisons test. # p < 0.05, ### p < 0.01, and ### p < 0.001.
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<t>RORα</t> mediates n-3 PUFA-regulated rhythmic oscillations in molecular biology (A and B) Optimal conformation of the <t>RORα</t> <t>(PDB</t> ID: 4s15 ) docking complex bound with DHA (A) and EPA (B). The RORα structure is shown as ribbon in blue while in light blue for the enlarged zoom of the active pocket. Detailed binding residues are shown as yellow sticks, and hydrophobic interactions are all shown as semitransparent gray spheres. The yellow dotted line is hydrogen bonding, measuring a distance range of 2.0–3.0 Å; a gray translucent surface indicates hydrophobic action. (C) Free energy of binding of 4s15 receptor to ligands (4DB, DHA-derivate MaR1, DHA, and EPA) was estimated with the highest score. Molecular docking estimates were performed by AutodockVina. (D–G) Effects of 100 μM DHA/EPA plus 300 μM PA on the oscillations of circadian clock genes including Clock (D), Bmal1 (E), Per2 (F), and Rev-erb (G), in RORα-KO hypothalamic neuron cells. The relative mRNA expressions were detected at ZT 0, 4, 8, 12, 16, 20, 24, 28, and 32 after synchronization, and oscillation curves were fitted by Cosinor algorithm ( n = 3 for each group, biological replicates). (H–K) Representative immunoblotting of clock proteins including (H and J) CLOCK and (I and K) REV-ERB at ZT 4, 12, and 20 after synchronization in RORα-KO hypothalamic neuron cells ( n = 3 for each group, biological replicates). For (D)–(K), data were expressed as mean ± SEM. Statistical differences between PA treatment versus other groups at same time point were determined by one-way ANOVA followed by the Dunnett’s multiple comparisons test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Statistical differences between the same group at different time points; statistical differences were determined by one-way ANOVA followed by the Tukey’s multiple comparisons test. # p < 0.05, ### p < 0.01, and ### p < 0.001.
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RORα mediates n-3 PUFA-regulated rhythmic oscillations in molecular biology (A and B) Optimal conformation of the RORα (PDB ID: 4s15 ) docking complex bound with DHA (A) and EPA (B). The RORα structure is shown as ribbon in blue while in light blue for the enlarged zoom of the active pocket. Detailed binding residues are shown as yellow sticks, and hydrophobic interactions are all shown as semitransparent gray spheres. The yellow dotted line is hydrogen bonding, measuring a distance range of 2.0–3.0 Å; a gray translucent surface indicates hydrophobic action. (C) Free energy of binding of 4s15 receptor to ligands (4DB, DHA-derivate MaR1, DHA, and EPA) was estimated with the highest score. Molecular docking estimates were performed by AutodockVina. (D–G) Effects of 100 μM DHA/EPA plus 300 μM PA on the oscillations of circadian clock genes including Clock (D), Bmal1 (E), Per2 (F), and Rev-erb (G), in RORα-KO hypothalamic neuron cells. The relative mRNA expressions were detected at ZT 0, 4, 8, 12, 16, 20, 24, 28, and 32 after synchronization, and oscillation curves were fitted by Cosinor algorithm ( n = 3 for each group, biological replicates). (H–K) Representative immunoblotting of clock proteins including (H and J) CLOCK and (I and K) REV-ERB at ZT 4, 12, and 20 after synchronization in RORα-KO hypothalamic neuron cells ( n = 3 for each group, biological replicates). For (D)–(K), data were expressed as mean ± SEM. Statistical differences between PA treatment versus other groups at same time point were determined by one-way ANOVA followed by the Dunnett’s multiple comparisons test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Statistical differences between the same group at different time points; statistical differences were determined by one-way ANOVA followed by the Tukey’s multiple comparisons test. # p < 0.05, ### p < 0.01, and ### p < 0.001.

Journal: Cell Reports Medicine

Article Title: Marine n-3 polyunsaturated fatty acids slow sleep impairment progression by regulating central circadian rhythms in type 2 diabetes

doi: 10.1016/j.xcrm.2025.102128

Figure Lengend Snippet: RORα mediates n-3 PUFA-regulated rhythmic oscillations in molecular biology (A and B) Optimal conformation of the RORα (PDB ID: 4s15 ) docking complex bound with DHA (A) and EPA (B). The RORα structure is shown as ribbon in blue while in light blue for the enlarged zoom of the active pocket. Detailed binding residues are shown as yellow sticks, and hydrophobic interactions are all shown as semitransparent gray spheres. The yellow dotted line is hydrogen bonding, measuring a distance range of 2.0–3.0 Å; a gray translucent surface indicates hydrophobic action. (C) Free energy of binding of 4s15 receptor to ligands (4DB, DHA-derivate MaR1, DHA, and EPA) was estimated with the highest score. Molecular docking estimates were performed by AutodockVina. (D–G) Effects of 100 μM DHA/EPA plus 300 μM PA on the oscillations of circadian clock genes including Clock (D), Bmal1 (E), Per2 (F), and Rev-erb (G), in RORα-KO hypothalamic neuron cells. The relative mRNA expressions were detected at ZT 0, 4, 8, 12, 16, 20, 24, 28, and 32 after synchronization, and oscillation curves were fitted by Cosinor algorithm ( n = 3 for each group, biological replicates). (H–K) Representative immunoblotting of clock proteins including (H and J) CLOCK and (I and K) REV-ERB at ZT 4, 12, and 20 after synchronization in RORα-KO hypothalamic neuron cells ( n = 3 for each group, biological replicates). For (D)–(K), data were expressed as mean ± SEM. Statistical differences between PA treatment versus other groups at same time point were determined by one-way ANOVA followed by the Dunnett’s multiple comparisons test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Statistical differences between the same group at different time points; statistical differences were determined by one-way ANOVA followed by the Tukey’s multiple comparisons test. # p < 0.05, ### p < 0.01, and ### p < 0.001.

Article Snippet: In order to investigate the interaction between DHA/EPA and mouse RORα (PDB ID: 4s15 ), we obtained the crystal structure of RORα and its original ligand (3beta,4alpha,5beta,14beta)-3-hydroxy-4-methylcholesta-8,24-diene-4-carboxylic acid (4D8) from the RCSB Protein DataBank ( http://www.rcsb.org/ ).

Techniques: Binding Assay, Western Blot