Fig. 3

Adipsin and Irak2 bind directly in cardiomyocytes. a Schematic representation of the experimental protocol. The cardiomyocytes lysis prey protein was obtained by immobilizing GST-Adipsin with equilibrated glutathione agarose. Then, the eluted protein complexes were employed for mass spectrometry analysis and subsequent immunoblotting. b Proteins bound and pulled down by purified GST-Adipsin in cardiomyocytes. c, d Co-IP results indicating the interaction between Adipsin and Irak2 following the infection with Ad-Adipsin. e Co-IP results indicating the interaction between Adipsin and Irak2 with or without PA challenge following the infection with Ad-Adipsin (n = 5). f Colocalization images and analysis of Adipsin (green) and Irak2 (red) proteins in cardiomyocytes. Scale bar = 50 μm. Statistical significance was determined using a two-tailed Student’s t test. All data are represented with mean ± SD. ^*P < 0.05 vs. Control. LC–MS/MS liquid chromatography-tandem mass spectrometry, Co-IP Co-immunoprecipitation, DAPI 4′,6-diamidino-2-phenylindole, Irak2 interleukin-1 receptor-associated kinase-like 2, PA palmitate, GST glutathione-S-transferase, IB immunoblotting