Journal: Scientific Reports

Article Title: M2 macrophages undergoing ferroptosis regulate uterine smooth muscle contraction to facilitate labor onset

doi: 10.1038/s41598-025-33523-4

Figure Lengend Snippet: Ferroptosis-related genes and pathways enriched in the transcriptomic profile of uterine myometrium during labor onset. ( A ) Box plots showing the expression profiles before and after normalization. ( B ) Venn diagram of differentially expressed genes (DEGs) from two GEO datasets ( GSE163773 and GSE186763 ) and ferroptosis-related genes (FRGs) from the FerrDb V2 database. ( C ) Heatmap of 49 ferroptosis-related differential genes (FRDGs) intersected from DEGs and FRGs. ( D ) Pathway enrichment analysis using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database of 49 FRDGs. ( E ) Functional clustering of FRDGs based on the Gene Ontology (GO) database.

Article Snippet: The cells were treated with 10 μM Ferrostatin-1 (Fer-1, a ferroptosis inhibitor, S7243, Selleck Chemicals, USA) and 1 μM (1 S,3R)-RSL3 (RSL3, a ferroptosis activator, S8155, Selleck Chemicals, USA) to establish a ferroptosis model . THP-1 (Tohoku Hospital Pediatrics-1, Procell, Wuhan, China), a human monocytic leukemia cell line, was maintained in RPMI1640 medium (MeilunBio, Dalian, China) containing 10% FBS (Procell, Wuhan, China) and 1% penicillin-streptomycin (Procell, Wuhan, China).

Techniques: Expressing, Functional Assay

Journal: Scientific Reports

Article Title: M2 macrophages undergoing ferroptosis regulate uterine smooth muscle contraction to facilitate labor onset

doi: 10.1038/s41598-025-33523-4

Figure Lengend Snippet: Ferroptosis-related proteins construct a protein–protein interaction network and diagnostic model for the onset of labor. ( A ) The protein-protein interaction (PPI) network analysis on the 49 FRDGs using the String database. ( B ) Calculating the number of relationship pairs between node genes of PPI network. ( C ) A volcano plot of the differential expression of 23 key FRGs. ( D ) The interaction network by using Cytoscape software. ( E ) 16 FRDGs were identified significantly differentially expressed in the validation dataset GSE181348 (TNL, n = 10; TL, n = 15). ( F ) Receiver operating characteristic (ROC) curve analysis for 12 key genes. ( G ) Constructing a diagnostic model for labor onset in the uterine myometrium.

Article Snippet: The cells were treated with 10 μM Ferrostatin-1 (Fer-1, a ferroptosis inhibitor, S7243, Selleck Chemicals, USA) and 1 μM (1 S,3R)-RSL3 (RSL3, a ferroptosis activator, S8155, Selleck Chemicals, USA) to establish a ferroptosis model . THP-1 (Tohoku Hospital Pediatrics-1, Procell, Wuhan, China), a human monocytic leukemia cell line, was maintained in RPMI1640 medium (MeilunBio, Dalian, China) containing 10% FBS (Procell, Wuhan, China) and 1% penicillin-streptomycin (Procell, Wuhan, China).

Techniques: Construct, Diagnostic Assay, Quantitative Proteomics, Software, Biomarker Discovery

Journal: Scientific Reports

Article Title: M2 macrophages undergoing ferroptosis regulate uterine smooth muscle contraction to facilitate labor onset

doi: 10.1038/s41598-025-33523-4

Figure Lengend Snippet: Correlation between core immune cells and key ferroptosis-related genes during labor onset. ( A ) The correlation between core immune cells and key FRDGs in GSE163773 and GSE186763 by corrplot R package. ( B ) HMOX1 showed a correlation with M2 macrophages in TL and TNL groups by Spearman. ( C ) The correlation between HMOX1 and key FRDGs in GSE163773 and GSE186763 by Spearman.

Article Snippet: The cells were treated with 10 μM Ferrostatin-1 (Fer-1, a ferroptosis inhibitor, S7243, Selleck Chemicals, USA) and 1 μM (1 S,3R)-RSL3 (RSL3, a ferroptosis activator, S8155, Selleck Chemicals, USA) to establish a ferroptosis model . THP-1 (Tohoku Hospital Pediatrics-1, Procell, Wuhan, China), a human monocytic leukemia cell line, was maintained in RPMI1640 medium (MeilunBio, Dalian, China) containing 10% FBS (Procell, Wuhan, China) and 1% penicillin-streptomycin (Procell, Wuhan, China).

Techniques:

Journal: Scientific Reports

Article Title: M2 macrophages undergoing ferroptosis regulate uterine smooth muscle contraction to facilitate labor onset

doi: 10.1038/s41598-025-33523-4

Figure Lengend Snippet: Ferroptosis levels upregulation in human uterine smooth muscle tissue during labor onset and regulate phenotypic changes in HUSMCs. ( A ) The mRNA levels of key ferroptosis genes by RT-qPCR (HMOX1, TFRC, ACSL4, SLC39A14, NRF2, SLC40A1 and GPX4) in human myometrium of TNL and TL groups (TNL, n = 15; TL, n = 15). ( B ) The protein levels of key ferroptosis genes by WB (HMOX1, TFRC, ACSL4, SLC40A1 and GPX4) in human myometrium of TNL and TL groups. Original blots are presented in Supplementary Figure S5-S9. ( C )-( E ) The levels of Fe²⁺, T-GSH, and MDA in human myometrium of TNL and TL groups (TNL, n = 15; TL, n = 15). ( F ) The expression of the key gap junction protein CX43 in HUSMCs treated with DMSO, Ferrostatin-1 (Fer-1) and RSL3 ( n = 3 in every group). Original blots are presented in Supplementary Figure S10. ( G )-( H ) Phalloidin staining and statistics of F-actin (red) in HUSMCs treated with DMSO, Fer-1 and RSL3 ( n = 3 in every group). Scale bar: 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.001.

Article Snippet: The cells were treated with 10 μM Ferrostatin-1 (Fer-1, a ferroptosis inhibitor, S7243, Selleck Chemicals, USA) and 1 μM (1 S,3R)-RSL3 (RSL3, a ferroptosis activator, S8155, Selleck Chemicals, USA) to establish a ferroptosis model . THP-1 (Tohoku Hospital Pediatrics-1, Procell, Wuhan, China), a human monocytic leukemia cell line, was maintained in RPMI1640 medium (MeilunBio, Dalian, China) containing 10% FBS (Procell, Wuhan, China) and 1% penicillin-streptomycin (Procell, Wuhan, China).

Techniques: Quantitative RT-PCR, Expressing, Staining

Journal: Scientific Reports

Article Title: M2 macrophages undergoing ferroptosis regulate uterine smooth muscle contraction to facilitate labor onset

doi: 10.1038/s41598-025-33523-4

Figure Lengend Snippet: Ferroptosis-driven M2 macrophages secrete proinflammatory cytokines to modulate uterine smooth muscle contraction and facilitate labor onset. The figure was generated by Figdraw.

Article Snippet: The cells were treated with 10 μM Ferrostatin-1 (Fer-1, a ferroptosis inhibitor, S7243, Selleck Chemicals, USA) and 1 μM (1 S,3R)-RSL3 (RSL3, a ferroptosis activator, S8155, Selleck Chemicals, USA) to establish a ferroptosis model . THP-1 (Tohoku Hospital Pediatrics-1, Procell, Wuhan, China), a human monocytic leukemia cell line, was maintained in RPMI1640 medium (MeilunBio, Dalian, China) containing 10% FBS (Procell, Wuhan, China) and 1% penicillin-streptomycin (Procell, Wuhan, China).

Techniques: Generated