Journal: Poultry Science

Article Title: Synergistic effect of genistein and adiponectin reduces fat deposition in chicken hepatocytes by activating the ERβ-mediated SIRT1-AMPK signaling pathway

doi: 10.1016/j.psj.2024.103734

Figure Lengend Snippet: Effect of genistein on the protein levels of estrogen receptors and Erk-PPARγ signaling pathway related factors in chicken adipocytes. (A) Immunoblot of ERα, ERβ, GPER1, and Tublin β protein; (B) The protein levels of ERα, ERβ, and GPER1; (C) Immunoblot of p-Erk, t-Erk, PPARγ, and Tublin β protein; (D) The protein levels of p-Erk/t-Erk and PPARγ. Data are expressed as means ± SEM (n = 3). ** P < 0.01, compared with the control group.

Article Snippet: After that, the membranes were incubated overnight with the rabbit antibodies against GPER1 (Cat#A10217; 1:1000 dilution; ABclonal Technology, Wuhan, China), Nampt (Cat#A0256; 1:1500 dilution; ABclonal Technology, Wuhan, China), ERβ (Cat#A2546; 1:1500 dilution; ABclonal Technology, Wuhan, China), PPARγ (Cat#A19676; 1:1500 dilution; ABclonal Technology, Wuhan, China), ACTB (Cat#AC026; 1:20000 dilution; ABclonal Technology, Wuhan, China), ERα (Cat#8644; 1:2000 dilution; Cell Signaling Technology, Boston, USA), Erk (Cat#4695; 1:2000 dilution; Cell Signaling Technology, Boston, MA), p-Erk (Cat#4370; 1:2000 dilution; Cell Signaling Technology), AMPK (Cat#5831; 1:2000 dilution; Cell Signaling Technology), p-AMPK (Cat#2535; 1:1000 dilution; Cell Signaling Technology), ACC (Cat#3662; 1:2000 dilution; Cell Signaling Technology), p-ACC (Cat#3661; 1:1000 dilution; Cell Signaling Technology) and SIRT1 (Cat#BS6494; 1:1000 dilution; Bioworld Technology, Nanjing, China).

Techniques: Western Blot, Control

Journal: Poultry Science

Article Title: Synergistic effect of genistein and adiponectin reduces fat deposition in chicken hepatocytes by activating the ERβ-mediated SIRT1-AMPK signaling pathway

doi: 10.1016/j.psj.2024.103734

Figure Lengend Snippet: Genistein promotes adiponectin secretion through binding with ERβ in adipocytes. ICP-1 cells were pretreated with vehicle, GPER1 inhibitor G15 (1 μM; 12h), GPER1 activator G1 (1 μM; 1h), ER antagonist Fulvestrant (1 μM; 12h) or ERβ antagonist PHTPP (10 μM; 1h); and then the cells were treated with 0 or 40 μM GEN for another 24 h. (A) Immunoblot of p-Erk, t-Erk, PPARγ, and Tublin β protein; (B–C) The protein levels of p-Erk/t-Erk and PPARγ; (D) APN mRNA expression; (E) Immunoblot of p-Erk, t-Erk, PPARγ, and Tublin β protein; (F–G) The protein levels of p-Erk/t-Erk and PPARγ; (H) APN mRNA expression. (I) Immunoblot of p-Erk, t-Erk, PPARγ, and Tublin β protein; (J–K) The protein levels of p-Erk/t-Erk and PPARγ; L: APN mRNA expression; Data are expressed as means ± SEM (n = 3). ** P < 0.01, comparison with the respective control groups; NS, no significant difference between the indicated groups.

Article Snippet: After that, the membranes were incubated overnight with the rabbit antibodies against GPER1 (Cat#A10217; 1:1000 dilution; ABclonal Technology, Wuhan, China), Nampt (Cat#A0256; 1:1500 dilution; ABclonal Technology, Wuhan, China), ERβ (Cat#A2546; 1:1500 dilution; ABclonal Technology, Wuhan, China), PPARγ (Cat#A19676; 1:1500 dilution; ABclonal Technology, Wuhan, China), ACTB (Cat#AC026; 1:20000 dilution; ABclonal Technology, Wuhan, China), ERα (Cat#8644; 1:2000 dilution; Cell Signaling Technology, Boston, USA), Erk (Cat#4695; 1:2000 dilution; Cell Signaling Technology, Boston, MA), p-Erk (Cat#4370; 1:2000 dilution; Cell Signaling Technology), AMPK (Cat#5831; 1:2000 dilution; Cell Signaling Technology), p-AMPK (Cat#2535; 1:1000 dilution; Cell Signaling Technology), ACC (Cat#3662; 1:2000 dilution; Cell Signaling Technology), p-ACC (Cat#3661; 1:1000 dilution; Cell Signaling Technology) and SIRT1 (Cat#BS6494; 1:1000 dilution; Bioworld Technology, Nanjing, China).

Techniques: Binding Assay, Western Blot, Expressing, Comparison, Control

Journal: Neuron

Article Title: Pathogenic DDX3X mutations impair RNA metabolism and neurogenesis during fetal cortical development

doi: 10.1016/j.neuron.2020.01.042

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Alexa647-alkyne , Thermo Fisher Scientific , A10278.

Techniques: Western Blot, Virus, Recombinant, Fluorescence, Luciferase, Synthesized, ATPase Assay, Negative Control, In Situ Hybridization, Plasmid Preparation, Protein Purification, Software

Journal: Nucleic Acids Research

Article Title: Detection of base analogs incorporated during DNA replication by nanopore sequencing

doi: 10.1093/nar/gkaa517

Figure Lengend Snippet: Design and assembly of training templates with single and multiple modifications at defined locations. ( A ) A schematic of the two-step assembly process of modified training templates. The first step is nicking the vector with the restriction enzyme nb.BbvCI and the second- ligation of a ssDNA oligo which carries 1, 2 or 14 modifications. The vector is then linearized with SpeI-HF to generate a 6184bp modified template, sequenced on the MinION. ( B ) Sequence of the single, double and multiple modification oligos used for ligation. ( C ) Dot blots with ligation products for single modification templates, carrying AF647, AF488, Biotin or a BrdU moiety, demonstrating efficient replacement with the modified oligo. ( D ) Dot blots with ligation products for multi-modified templates, carrying 14 CldU or 14 IdU modifications. ( E ) A schematic illustrating the sequencing of a training template with one or more modifications through a nanopore.

Article Snippet: Separate reactions were carried out with the following azides: sodium azide (Sigma S2002), nicotinoyl azide (Sigma CDS006775), 6-azido-6-deoxy- d -glucose (Sigma 712760) Biotin azide (Thermo Fisher B10184), AF488 azide (Thermo Fisher A10266), AF647 azide (Thermo Fisher A10277) and a ssDNA oligo purchased from Integrated DNA Technologies: 5′-GGATAGCCTC/3AzideN/-3′.

Techniques: Modification, Plasmid Preparation, Ligation, Sequencing

Journal: Nucleic Acids Research

Article Title: Detection of base analogs incorporated during DNA replication by nanopore sequencing

doi: 10.1093/nar/gkaa517

Figure Lengend Snippet: Detecting a series of modifications with increasing molecular weight through MinION sequencing. A violin plot of the known modification site and the surrounding bases for ( A ) EdU, ( B ) CldU, ( C ) BrdU, ( D ) azide, ( E ) IdU, ( F ) nicotine, ( G ) glucose, ( H ) biotin, ( I ) streptavidin purified biotin, ( J ) AF488, ( K ) AF647, ( L ) 10 bp ssDNA oligo, attached to position 3073 by click cycloaddition and ( M ) dual-labeled template with Biotin at 3049 and IdU at 3098. In each panel, the first line denotes position, followed by the base, the P -value for a Kolmogorov–Smirnov test and the P -value for Stouffer's method. The second line shows a violin plot of mean normalized signal from the control and modified sample, the third line contains the logarithm of the P -value (pv) for Kolmogorov–Smirnov test, and the fourth line displays the logarithm of the combined P -value, calculated using Stouffer's method. ‘DS 1’ in blue represents the non-modified oligo sample, while ‘DS 2’ in green stands for the modified template; ‘- strand’ denotes reverse strand. Black stars represent unmodified T’s, while a modified position is denoted by a red star. B = biotin and I = IdU.

Article Snippet: Separate reactions were carried out with the following azides: sodium azide (Sigma S2002), nicotinoyl azide (Sigma CDS006775), 6-azido-6-deoxy- d -glucose (Sigma 712760) Biotin azide (Thermo Fisher B10184), AF488 azide (Thermo Fisher A10266), AF647 azide (Thermo Fisher A10277) and a ssDNA oligo purchased from Integrated DNA Technologies: 5′-GGATAGCCTC/3AzideN/-3′.

Techniques: Molecular Weight, Sequencing, Modification, Purification, Labeling, Control