Fig 1: Serum myonectin levels were correlated with the severity and pancreatic necrosis of AP patients. (A) Flowchart of the patients with AP in the clinical study. (B) ELISA analysis of serum Myonectin levels in the healthy control group and different severity levels of AP patients. (C) Correlation analysis of patient serum Myonectin with ANC and OF. (D) Correlation analysis of patient serum Myonectin with CTSI. (E) Correlation coefficient heat map of serum Myonectin levels upon admission with other clinical parameters. ns p ≥ 0.05, * p<0.05, **p<0.01, ***p<0.001. HC, health check; MAP, mild AP; MSAP, moderately severe AP; SAP, severe AP; ANC, absolute neutrophil count; OF, organ failure; CTSI, CT severity index; WBC, white blood cells; PCT, procalcitonin; CRP, C-reactive protein; HCT, hematocrit; LDH, lactate dehydrogenase
Fig 2: Significant overexpression of the myokines myonectin in SAP mice. (A, B) Western blot and quantitative analysis of Myonectin in muscle tissue at different time points post PDL surgery. (C) qPCR detection of Myonectin in muscle tissue at different time points post PDL surgery. (D) ELISA detection of serum Myonectin levels in mice at different time points post PDL surgery. N ≥ 4, ns p ≥ 0.05, * p<0.05, **p<0.01, ***p<0.001
Fig 3: RNA-seq data identified pathways related to iron metabolism and ferroptosis as targets of myonectin. (A) Myonectin related genes were screened in the String database. (B) Overlap of myonectin-related genes from the String database with RNA-seq data in the SAP model identified 17 common genes. (C) KEGG enrichment analysis of these 17 genes. (D) GO enrichment analysis of these 17 genes
Fig 4: Myonectin induced iron accumulation and intensified acinar cell ferroptosis. (A) Immunofluorescence imaging of pancreatic acinar cells stained with FerroOrange (1200x). (B) Quantitative analysis of fluorescence intensity (N ≥ 5). (C) Western blot detection of key ferroptosis protein GPX4 expression levels in PACs. (D) Quantitative analysis of GPX4 expression changes. (E) C11-BODIPY measurement of cellular lipid peroxidation levels. (F) MDA assessment of lipid peroxidation levels in pancreatic tissue. N ≥ 4, ns p ≥ 0.05, * p<0.05, **p<0.01, ***p<0.001
Fig 5: Exogenous administration of recombinant myonectin protein aggravates acinar cell necrosis. (A) PACs were treated with CCK, together with gradient doses of Myonectin for 6 h; the levels of LDH release are shown. (B) Immunofluorescence imaging of pancreatic acinar cells stained with Calcein-AM/PI (1200x). (C) Representative images of pancreatic tissue H&E staining in PDL mice after treatment with gradient doses of recombinant Myonectin protein. (D) Pathological scoring of pancreatic tissue (edema, necrosis, inflammatory cell infiltration). (E) Serum amylase levels. (F) Serum lipase levels. CCK, cholecystokinin; AM, acetoxymethyl, represents living cells; PI, propidium iodide, represents dead cells. N ≥ 5, ns p ≥ 0.05, * p<0.05, **p<0.01, ***p<0.001
Supplier Page from CUSABIO Technology LLC for Human Protein FAM132B(FAM132B) ELISA Kit