Figure 5
From: Metabolomics and the pig model reveal aberrant cardiac energy metabolism in metabolic syndrome
Fumarase O-GlcNAcylation and activity response to MS and LD. (a) The bar diagram shows the precursor fumarate (FUM) to malate (MAL) product ratios in LD and MS. The FUM and MAL content is determined based on the LC/MS-MS data in LD (n = 8) and MS pig (n = 7). The FUM content is taken as a base (100%) to calculate the MAL content in a FUM: MAL analysis (454.8 ± 67.09%) in LD and (159.5 ± 27.81%) in MS. The product data are presented as means ± SEM; **p < 0.01, MS (n = 8), LD (n = 7): the precursor and product identities are shown at the bottom of the diagram. (b) FH activity was measured in total tissue lysates (10 μg protein/reaction LD n = 4, MS n = 4) at 37 °C. All activity measurements were conducted in duplicate at 37 oC for 20 min, and data are presented as means ± SD; ****p = 0.0001 in relative units/µg protein (U/ µg protein). (c) The diagram summarizes the western blot data with the WGA affinity capture of O-GlcNAcylated FH in total protein lysates from MS (n = 4) and LD (n = 4): specific immunological reaction (pixel density) for FH in the input fractions -IN, flow through- F, WGA bound – B, bound to control beads without WGA- C. As a base for comparison is used the FH-positive reaction in the input fraction (100%). Data are represented as means ± SD; ***p < 0.001.
