Fig. 5

Kinome profiling of unaffected mammary glands revealed regulation of PKD1-PKC-α-PKA-MEK3 and AMPK by diet exposure. To profile the activated kinome, unaffected mammary glands from mice at 15 weeks of age were collected for multiplexed inhibitor bead (MIB) affinity chromatography and mass spectrometry analysis. a MIBs consist of mixtures of Sepharose beads with covalently immobilized, linker-adapted, broad pan-kinase inhibitors (listed on left of column) are designed to capture kinases in the active state in a reproducible and reliable assay. Two to four samples were pooled into a total of 3 runs per diet group. (N = 8 mice in each diet group). All kinase activity is normalized to 10 %-fed controls. b Mean kinase activity is presented to compare kinases in the unaffected mammary glands that were significantly different between mice in the 60 % group and 60–10 % group. *P < 0.005, ^P < 0.05, 60 % vs. 60–10 %) In b, no error bar is present in pooled samples when kinases were down-regulated below level of detection and only 1 run detected activity. c, d Protein–protein interactions of significantly altered kinases in unaffected mammary gland of mice on 60–10 % diet compared to mice on 60 % diet. c Search Tool for the Retrieval of Interacting Genes/Proteins (STRING version 10) was used to visualize known protein–protein interactions between significantly regulated kinases. Confidence view was shown. Stronger associations are represented by thicker lines. d Cartoon of a subset of kinases regulated by HFD and reversed by weight loss and the contribution of obesity-induced leptin signaling