Fig 1: Annexin A1 production is enhanced during zymosan-induced inflammation. Mice were injected i.p. with zymosan (Zym) and peritoneal lavage supernatant, peritoneal macrophages (PM), and spleens were collected at 6, 24, or 72 h after Zym injection. (A,B) The levels of Annexin A1 (AnxA1) and TNF-a-stimulated gene-6 (TSG6) mRNA over time in PM and spleens analyzed by real-time PCR and normalized to that of hypoxanthine guanine phosphoribosyl transferase (Hprt) mRNA. (C,D) The abundance of AnxA1 and TSG6 in peritoneal lavage fluid (PLF) as assessed by ELISA. (E) Immunoblots analysis of AnxA1 in spleen homogenates. Lower pannel: Densitometric analysis of AnxA1 normalized to that of ß-actin. Values represent the means ± SEM of results from five (A,B) or three mice (C–E). * p < 0.05, *** p < 0.001 compared with control at a given time point (Student’s t-test).
Fig 2: Annexin A1 enhances STAT6 activation and PPAR? expression and activation in BMDM and peritoneal macrophages. Immunoblots analysis of the relative amounts of phosphorylated STAT6/total STAT6 in mouse BMDM (A) and peritoneal macrophages (F) stimulated with Annexin A1 (AnxA1) for 2 h. Densitometric analysis of the relative abundance of the indicated proteins normalized to that of STAT6. Left: Immunofluorescence staining of phosphorylated STAT6 (green) in BMDM (B) and peritoneal macrophages (G) after 100 ng/mL AnxA1 treatment. The imaging medium was Vectashield fluorescence mounting medium containing DAPI. Original magnification: ×400. Scale bars = 40 µm. Representative results from three independent experiments are shown. Right: The ratio of phospho-STAT6 (+) over DAPI (+) macrophages is presented in the bar graph. Immunoblots analysis of the relative amounts of PPAR?, CD36, and ß-actin in BMDM (C,E) and peritoneal macrophages (H,J) stimulated with AnxA1 for 24 h. Densitometric analysis of the relative abundance of the indicated proteins normalized to that of ß-actin. PPAR? activity in nuclear extracts from BMDM (D) and peritoneal macrophages (I) was analyzed at 24 h after AnxA1 treatment as described in the Methods. Values represent the mean ± SEM of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with control (Student’s t-test).
Supplier Page from Abcam for Recombinant Mouse Annexin A1/ANXA1 protein