Journal: Nature Metabolism
Article Title: Intestinal TM6SF2 protects against metabolic dysfunction-associated steatohepatitis through the gut–liver axis
doi: 10.1038/s42255-024-01177-7
Figure Lengend Snippet: a , Experimental schematic of untargeted metabolomic profiling on the culture supernatant of IECs isolated from Tm6sf2 ΔIEC and Tm6sf2 fl mice. Created with BioRender.com . b , PCA analysis and heat map of differential metabolites secreted by IECs from mice ( n = 8 per group). c , Metabolomics targeting free fatty acids on the cultural supernatant of isolated IECs ( n = 8 per group). d , Representative small intestine images of H&E staining and intestinal triglyceride of Tm6sf2 ΔIEC mice fed with NC ( n = 5 per group) or CD-HFD ( n = 9 per group). e , Silver staining coupled with mass spectrometry analysis after pull-down assay on mouse intestinal tissues. f , Co-immunoprecipitation of TM6SF2 and FABP5 using mouse intestinal proteins. g , MST assay for direct binding between TM6SF2 and FABP5 with dissociation constant ( K d ) provided. h , Representative structure of molecular docking between TM6SF2 (purple) and FABP5 (blue). i , Metabolomics targeting free fatty acids in TM6SF2 KO Caco2 cells with or without FABP5 overexpression ( n = 6 per group). j , Experimental schematic and intestinal permeability by FITC-labelled dextran assay in mice supplemented with the top differential free fatty acids ( n = 5 per group). k , Permeability of Caco2 cell monolayers treated with arachidic acid or arachidonic acid ( n = 3 per group). l , m , Heat map of differential faecal microorganisms ( l ) and LPA-targeting metabolomics on portal vein serum ( m ) of mice treated with free fatty acids ( n = 4–5 per group). Results are presented as the mean ± s.d. Experiments were repeated three times with similar results ( e and f ). Statistical significance was determined by two-tailed Student’s t -test ( c , d , i and k ) or one-way ANOVA followed by Turkey’s multiple comparison ( j and m ).
Article Snippet: The protein mixture was also incubated overnight at 4 °C with either anti-TM6SF2 antibody or anti-FABP5 antibody (RRID: AB_2100341, 12348-1-AP, Proteintech).
Techniques: Isolation, Staining, Silver Staining, Mass Spectrometry, Pull Down Assay, Immunoprecipitation, Binding Assay, Over Expression, Permeability, Two Tailed Test, Comparison