secretory factorswerequantifiedusing humanlysozymeelisakit  (Cusabio)

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

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 Schematic model of stem cell maintenance by Paneth cells in ISC niche. The solid arrows indicate activation and the blunt-ended arrows represent inhibition. b Intracellular cADPR in the sorting Paneth cells using an ELISA kit. Amounts of cADPR were normalized to total protein ( n = 3 biologically independent mice per group). c Endogenous cADPR levels in organoids treated with rapamycin, ALA, and MHY1485 were measured using an ELISA kit ( n = 3 biologically independent mice per group). d – i Crypts isolated from old mice were cultured with rapamycin, MHY1485, cADPR and 8-Br-cADPR together with/without ALA. Representative images (scale bars, 250 μm) of mouse intestinal organoids ( d ), quantification of organoid amount ( n = 7 biologically independent mice per group) ( e ), representative images (scale bars, 50 μm) of organoid buds ( f ), quantification of bud per organoid ( n = 13 organoids/group, obtained from 7 biologically independent mice) ( g ), representative staining (scale bars, 50μm) of EdU cells ( h ), quantification of EdU + cell amount in mouse intestinal organoids( i ). ( n = 10 organoids/group, obtained from 7 biologically independent mice). j Schematic models of ALA functions in Paneth cells to prevent ISC aging. Data are displayed as mean ± SD. One-way ANOVA analysis followed by Tukey’s multiple comparisons test was used in ( b , c , e , g , 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: Activation Assay, Inhibition, Enzyme-linked Immunosorbent Assay, Isolation, Cell Culture, Staining

Journal: Scientific Reports

Article Title: Azathioprine promotes intestinal epithelial cell differentiation into Paneth cells and alleviates ileal Crohn’s disease severity

doi: 10.1038/s41598-024-63730-4

Figure Lengend Snippet: Levels of Paneth cells markers in CD patients and hospitalized controls. ( a ) mRNA expression of LYZ , DEFA5 and DEFA6 in ileal samples from CD patients and hospitalized controls. ( b ) Microscopic images of IHC staining of LYZ in ileal samples from CD patients and healthy controls. Scale bar = 500, 20 µm (20x, 63x). ( c ) Intensity of the staining per crypt was measured using ImageJ software. ( d ) Lysozyme level was measured in fecal samples from CD patients treated or not treated with AZA by ELISA. ( e ) qPCR was performed to measure mRNA levels of KI67 in ileal biopsies CD patients and controls. ( f ) Representative images of ileal KI67 staining in CD patients and controls. Scale bar = 20 µm (63x). ( g ) Numbers of Ki67 + cells were counted using ImageJ software. ( a , c , e , g ) Kruskal–Wallis test; ( d ) Unpaired t test with Welch’s correction; results are shown as truncated violin plots with median represented as dashed line. * p < 0.05, ** p < 0.01. HC: hospitalized controls; CD: non-AZA treated CD patients; CD-A: azathioprine-treated CD patients; DEFA5 , 6: alpha-defensin 5, 6; LYZ : lysozyme.

Article Snippet: Lysozyme concentration in protein extracts from human stool samples (1:100) was measured using Human Lysozyme ELISA Kit (Abcam, ab267798, Cambridge, UK).

Techniques: Expressing, Immunohistochemistry, Staining, Software, Enzyme-linked Immunosorbent Assay