Ubinuclein Antibody - #DF8873

Figure 4 Decreased ATP5O-K51 crotonylation conduced to reduced ATP5O stability and decreased lipid metabolism. (a–c) Transfection of ATP5O-WT or K51A (inactive mutant) plasmid (in pcDNA3.1) into 293 T cells and western blot by ATP5O-K51 cr antibody showed that ATP5O-K51A had significantly decreased crotonylation level (a, b), accordingly, ATP5O K51A-transfected 293 T cells had significantly lower cytoplasmic ATP level than ATP5O-WT-transfected 293 T cells. (d–g) Western blot showed that inhibition of ATP5O crotonylation with a competitive peptide TAC (CYGRKKRRQRRRATALYSAASKEKKL) significantly reduced ATP5O-K51 crotonylation (d, e) and gross ATP5O (f, g) levels, similar to CS treatment. (h, i). A region of Ubinuclein (UBN) shared the highest similarity with TAC peptide (Supp. Figure 8); however, TAC treatment did not change UBN level at all. (j, k) Western blot showed that inhibition of ATP5O crotonylation with TAC significantly decreased STAT5A protein level. (l) TAC treatment significantly reduced liver ATP level. (m–o) Quantitative lipidome showed that at the threshold of ATP5O−TAC/Ctr≥1.5 or ≤0.67 , there were 174 differentially upregulated or downregulated lipid metabolites (DULMs or DDLMs), among this, 22.41% (39/174) was phospholipid (PE & PC & PI), and 82.05% (32/39) of phospholipid was downregulated. (p) Representative downregulated phospholipid in TAC-treated group. (q–s) Blood biochemical index assay showed that multiple indexes in TAC group, including CREA, SOD, and HDL-C, were similar to CS group and significantly different from control group. Tubulin or actin was used as a loading control.