Journal: Nature Communications
Article Title: Amelioration of colorectal cancer-associated fibroblasts in immunosuppressive microenvironment by ferroptosis-based nanotherapy
doi: 10.1038/s41467-026-69462-5
Figure Lengend Snippet: a Schematic illustration of cell viability and transwell migration assays using HCT116 cells treated with supernatants from NFs, CAFs, or NPs-treated CAFs. b Transwell migration images presenting the number of migrated HCT116 cells treated with CM from NFs, CAFs, or NPs-treated CAFs for 24 h. c Cell viability of HCT116 cells treated as in ( b ) by the CCK8 assay. d Western blot of E-cadherin, N-cadherin, and Vimentin expression in HCT116 cells treated with CM from NFs, CAFs, or NPs-treated CAFs for 24 h; β-actin as a loading control. e , f Bubble plots of ( e ) GO and ( f ) KEGG pathway enrichment analyses for differentially expressed genes (DEGs) in NPs-treated CAFs compared to controls. g , h MSigDB Hallmark GSEA analysis showing enrichment of ( g ) chemokine activity and ( h ) ferroptosis pathways in NPs-treated CAFs compared to controls. ES: enrichment score; NES: normalized enrichment score. i Heatmap and histogram displaying gene expression levels of DEGs (log₂|FC | ≥ 1) from three chemokine-related GO pathways (GO:0016493, GO:0048020, GO:0008009) in NPs-treated CAFs compared to controls, ranked by differential expression. j QRT-PCR quantification of CCL3 , CXCL12 , and CCR7 expression in NPs-treated CAFs compared to controls. k , l Anti-human ELISA quantification of ( k ) CCL3 and ( l ) CXCL12 concentrations in supernatants from CAFs treated without or with NPs for 24 h. m Western blot of MAPK, p-MAPK (Thr180/Tyr182), NF-κB, p-NF-κB (Ser536), AKT, and p-AKT (Ser473) expression in CAFs treated without or with NPs for 24 h; β-actin as a loading control. n = 3 independent experiments in ( b , c ) and ( j – l ). Western blot experiments in ( d , m ) are repeated independently three times with similar results. Data are presented as means ± SD. Statistical analyses are performed using one-way ANOVA with multiple comparisons for ( b , c ); and two-tailed unpaired Student’s t -test for ( j – l ). Source data are provided as a Source Data file.
Article Snippet: Human colonic epithelial cell line NCM460, human CRC cell line HCT116, murine CRC cell lines CT26 and MC38, the human monocytic leukemia cell line THP-1, and murine fibroblast cell lines (BALB/3T3 and NIH3T3) were originally obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and China Infrastructure of Cell Line Resources, Institute of Basic Medical Sciences, and Chinese Academy of Medical Sciences.
Techniques: Migration, CCK-8 Assay, Western Blot, Expressing, Control, Activity Assay, Gene Expression, Quantitative Proteomics, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Two Tailed Test
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