Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A Volcano plot showing differentially abundant metabolites in healthy infants ( n = 3 biologically independent samples) and NEC patients ( n = 6 biologically independent samples). B A lollipop plot displaying 15 upregulated and downregulated differential metabolites. C Analysis of endogenous L-cystine abundance in healthy infants and NEC patients. Data are presented as mean ± SD; * p < 0.05.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques:

Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A The standard chemical structure of L-cystine. B Cell viability measured by CCK-8 assay. C IL-6 mRNA and protein levels analyzed by qRT-PCR and Western blot. D IL-6 concentration in cell supernatant quantified by ELISA. E ROS generation assessed via flow cytometry. F Intracellular Fe²⁺ levels. G 4-HNE expression detected by immunofluorescence. Scale bar = 25 μm. H , I MDA, GSH, and SOD levels. J Western blot analysis of STAT3 phosphorylation and NCOA4/FTH1 protein levels. n = 3 biologically independent samples. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques: CCK-8 Assay, Quantitative RT-PCR, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Immunofluorescence, Phospho-proteomics

Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A Screening of IL-6 mRNA- and protein-binding targets using STARBASE and BioGRID databases. B RC3H1 protein levels post LPS and L-cystine treatment by Western blot. C Efficiency of RC3H1 knockdown and overexpression plasmids in NCM-460 cells. D IL-6 mRNA and protein levels after RC3H1 knockdown/overexpression by qRT-PCR and Western blot. E RIP-qPCR validation of RC3H1 binding to IL-6 mRNA. F IL-6 mRNA stability assay using Actinomycin D (5 μg/mL). G Co-IP analysis of RC3H1 and IL-6 interaction. H IL-6 and RC3H1 protein levels using Western blot. I IL-6 concentration in supernatant by ELISA. n = 3 biologically independent samples. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques: Protein Binding, Western Blot, Knockdown, Over Expression, Quantitative RT-PCR, Biomarker Discovery, Binding Assay, Stability Assay, Co-Immunoprecipitation Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Communications Biology

Article Title: L-cystine alleviates necrotizing enterocolitis by regulating ferroptosis and Th17 cell differentiation via the IL-6/STAT3 pathway

doi: 10.1038/s42003-025-09438-1

Figure Lengend Snippet: A Venn diagram of L-cystine and RC3H1 target interactions. B KIF11 and MMP2 protein levels post LPS and L-cystine treatment by Western blot. C DARTS assay confirming L-cystine binding to KIF11. D Efficiency of KIF11 knockdown and overexpression plasmids. E RC3H1 and IL-6 protein levels after KIF11 knockdown/overexpression by Western blot. F Co-IP analysis of KIF11 and RC3H1 interaction. G KIF11, IL-6, and RC3H1 protein levels. n = 3 biologically independent samples. Data are presented as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For therapeutic intervention, pups received daily oral L-cystine (200 mg/mL in saline solution; C6H12N2O4S2, 56-89-3, MCE, Monmouth Junction, NJ, USA) and intraperitoneal injections of the STAT3 inhibitor S3I-201 (0.5 mg/mL in DMSO and 20% cyclodextrin; 501919-59-1, MCE) twice weekly .

Techniques: Western Blot, Binding Assay, Knockdown, Over Expression, Co-Immunoprecipitation Assay