Journal: Nature Communications
Article Title: Lipoic acid functions in Paneth cells to prevent human intestinal stem cell aging
doi: 10.1038/s41467-025-61070-z
Figure Lengend Snippet: a , b Representative images of Paneth cells ( a , scale bars, 50 μm) and quantification in young and old human crypts ( b , n = 27 crypts/group from 6 independent donors). c – e Diagram of lysozyme distribution patterns: normal (D0), disordered (D1), diminished (D2), diffuse (D3), excluded (D4), enlarged (D5) ( c ). Representative staining showing lysozyme granules ( d , scale bars, 25 μm); white/yellow arrowheads indicate normal/abnormal granules. Quantification of distribution patterns ( e , n = 15 crypts/group from independent 6 donors). f – j Young and old mice were administered drinking water with vehicle or ALA (100 mg/kg/day) for 3 months. Representative images of Paneth cells ( f , scale bars, 75 μm), and quantification ( g , n = 33 crypts/group from 7 biologically independent mice), Representative images showing lysozyme granules ( h , scale bars, 10 μm; white/yellow arrowheads indicate normal/abnormal granules in Paneth cells). Distribution of lysozyme patterns ( i , n = 14 crypts/group from 7 mice). Representative TEM images of Paneth cells ( j , scale bars, 1μm; yellow/red stars indicate normal/abnormal granules). k , l Intestinal crypts from young and old humans were cultured with vehicle or ALA (100 μM) for 7 days. Representative lysozyme staining in organoids ( k , scale bars, 75μm) and Paneth cell quantification ( l , n = 7 organoids/group from 3 independent human donors). m – p Mouse intestinal crypts were cultured with vehicle or ALA (100 μM) for 6 days. Representative lysozyme staining ( m , scale bars, 75μm) and Paneth cell quantification ( n , n = 18 organoids/group from 7 mice). ELISA of Reg3g and Defa6 in organoid supernatants ( o , p , n = 3 biological replicates/group). All data are mean ± SD. P values were calculated by unpaired two-tailed t test ( b ), one-way ANOVA with Tukey’s test ( g , l , n , o , p ); for ( n ), p = 6e–9 (young vs old), 8e–5 (old vs old+ALA). Two-way ANOVA with Sidak’s test ( e ); for ( e ), p = 2e–8 (young vs old within D0 and D1–D5). Two-way ANOVA with Tukey’s test ( i ). Source data are provided as a Source Data file.
Article Snippet: Secretory factors were quantified using: Human Lysozyme ELISA Kit (CSB-E09135h, https://www.cusabio.com/ , CUSABIO)), Human DEFA6 ELISA Kit (JL29984, Jianglai biology, Shanghai), Human REG3γ ELISA Kit (ELK3316, ELK Biotechnology), Mouse DEFa6 ELISA Kit (JL29986, Jianglai Biology), Mouse REG3γ ELISA Kit (JM-11737M2, Jingmei Bio).
Techniques: Staining, Cell Culture, Enzyme-linked Immunosorbent Assay, Two Tailed Test