Journal: Frontiers in Pharmacology

Article Title: Polygonatum sibiricum combined with Poria cocos prevents spleen deficiency constipation via the gut microbiota–mediated brain–gut axis

doi: 10.3389/fphar.2026.1793441

Figure Lengend Snippet: Serum levels of 5-HT, CCK, VIP, and MTL in mice from each group. (A) 5-HT content; (B) CCK content; (C) MTL content; (D) VIP content. Note: NC: normal control group; NM: model group; HF: Polygonatum sibiricum and Poria cocos group.

Article Snippet: Mouse Serotonin (5-HT) ELISA Research Kit (Cat# JM-02726M2), Mouse Cholecystokinin (CCK) ELISA Research Kit (Cat# JM-02311M2), Mouse Motilin (MTL) ELISA Research Kit (Cat# JM-02775M2), Mouse Vasoactive Intestinal Peptide (VIP) ELISA Research Kit (Cat# JM-02729M2), the aforementioned kits were procured from Jiangsu Jingmei Biotechnology Co., Ltd. Superoxide Dismutase (SOD) Activity Assay Kit (BC0175), Malondialdehyde (MDA) Content Assay kit (BC0025), the aforementioned kits were procured from Beijing Solarbio Science & Technology Co., Ltd.

Techniques: Control

Journal: Cell Discovery

Article Title: TGM2-mediated serotonylation of GPX4 confers ferroptosis resistance to promote gastric tumorigenesis

doi: 10.1038/s41421-026-00885-6

Figure Lengend Snippet: a TGM2-dependent transamidation signals in recombinant GPX4 (monitored via monodansylcadaverine fluorescence) that were abolished by either cystamine inhibition or 5-HT competition. b Schematic illustration of the TGM2-dependent transamidation of glutamine residues in substrate proteins. c MS/MS spectra of the serotonylated GPX4 peptide (GPX4ser: Q55, Q77) derived from GPX4 (in vitro). The b ion refers to the N-terminal portion of the peptide, and the y ion indicates to the C-terminal portion. d SILAC experimental workflow for GPX4 serotonylation detection by LC-MS/MS using 5-HT and 5-HT- d 4 . TGM2 catalyzes the transamidation of glutamine residues in the GPX4 protein. e Chromatographic overlay of 5-HT and 5-HT- d 4 , demonstrating nearly identical retention times or 45.03 min and 45.05 min, respectively (top). MS analysis of 5-HT and 5-HT- d 4 revealed distinct isotopic peaks. Mass spectra of Light, with m/z of 727.36 and Heavy, with m/z of 729.37 m/z for 5-HT and 5-HT- d 4 , respectively, illustrating the clear separation between the light and heavy isotopic labels and confirming the use of deuterium-labeled serotonin for dual validation (bottom). f Tandem MS/MS spectrum of the peak at m/z 727.36 (5-HT Light) showing the fragmentation pattern of GPX4 at the serotonylated site, confirming the presence of 5-HT modification (top). Tandem MS/MS spectrum of the peak at m/z 729.37 (5-HT- d 4 Heavy) showing the corresponding fragmentation pattern, confirming the site-specific serotonylation of GPX4 (bottom). g Immunoblot and streptavidin pull-down analysis of Flag-GPX4 with various mutations (Q54A, Q77A, and Q2A) in HEK293T cells treated with 5-PT, confirming the loss of serotonylation at specific sites. Quantification is shown on the right (mean ± SEM; ordinary one-way ANOVA, n = 3).

Article Snippet: Each 50 μL reaction mixture contained 1× 50 mM HEPES reaction buffer (1× protease inhibitor), 5 mM 5-HT- d 4 (MCE, HY-B1473S) or 5 mM 5-HT combined with 5 mM CaCl 2 , 1.25 μg TGM2, and GPX4 recombinant protein (50 μg) and was established on ice.

Techniques: Recombinant, Fluorescence, Inhibition, Tandem Mass Spectroscopy, Derivative Assay, In Vitro, Multiplex sample analysis, Liquid Chromatography with Mass Spectroscopy, Labeling, Biomarker Discovery, Modification, Western Blot

Journal: Neuropsychopharmacology Reports

Article Title: Distinct Effects of Nonselective Rho‐Kinase Inhibitor Fasudil and Selective Rho‐Kinase 2 Inhibitor KD025 on Serotonin and Dopamine Release in the Nucleus Accumbens of Mice

doi: 10.1002/npr2.70124

Figure Lengend Snippet: Co‐expression of Rho‐kinases 1 and 2 with SERT or DAT. A: Representative immunoreactivity images of Rho‐kinase 1 (green), SERT (magenta), DAT (red), and Hoechst 33342 (blue) in the NAc (scale bar indicates 10 μm). B: Percentage of Rho‐kinase 1 and the SERT or DAT double‐positive area to the SERT or DAT‐positive area in the NAc. C: Representative immunoreactivity images of Rho‐kinase 2 (green), SERT (magenta), DAT (red), and Hoechst 33342 (blue) in the NAc (scale bar indicates 10 μm). D: Percentage of Rho‐kinase 2 and SERT or DAT double‐positive area relative to the SERT or DAT‐positive area in the NAc. Data represent the mean + SEM ( n = 5). DAT, dopamine transporter; SERT, serotonin transporter; NAc, nucleus accumbens.

Article Snippet: The following primary antibodies were used: anti‐Rho‐kinase 1 (Cat#21850–1‐AP, RRID:AB_10953526, 1:100 dilution; Proteintech, IL, USA), anti‐Rho‐kinase 2 (Cat# Ab125025 , RRID:AB_10972853, 1:200 dilution; Abcam, Cambridge, UK), anti‐serotonin transporter (SERT) (Cat#MSFR103270, 1:200 dilution; Nittobo Medical, Tokyo, Japan), and anti‐dopamine transporter (DAT) (Cat#sc‐32258, RRID:AB_627400, 1:250 dilution, Santa Cruz Biotechnology, Texas, USA).

Techniques: Expressing