Fig. 6

K. slooffiae promotes intestinal epithelial glycolysis via lysine desuccinylation induced by SIRT5. A, B Representative western blots of lysine-succinylated proteins and β-tubulin in IPEC-J2 cells using a pan-lysine succinylation antibody and an anti-β-tubulin antibody, respectively (Ctrl, Control; KS, K. slooffiae) (A). Quantification of lysine-succinylated proteins levels normalized to β-tubulin levels (B). C–E Glucose consumption (C), lactate production (D), and ATP content (E) in IPEC-J2 cells. F Volcano plot analysis of the identified proteins in IPEC-J2 cells. Up (red spots) indicates that the protein levels in KS group were higher than those in the Ctrl group. Down (blue spots) indicates that the protein levels in the KS group were lower than those in the Ctrl group. G Analysis of lysine succinyltransferases and lysine desuccinylases in protein levels. H Sirtuins (SIRTs) activity in IPEC-J2 cells. I, J Representative western blots of lysine-succinylated proteins and β-tubulin in IPEC-J2 cells using a pan-lysine succinylation antibody and an anti-β-tubulin antibody, respectively (I). Quantification of lysine-succinylated proteins levels normalized to β-tubulin levels (J). K–M Glucose consumption (K), lactate production (L), and ATP content (M) in IPEC-J2 cells. Data are presented as mean ± SEM and were assessed by one-way ANOVA with adjustment for multiple comparisons in B (n = 3), C–E (n = 6), J (n = 3), and K–M (n = 6). Data are presented as the mean ± SEM and were evaluated using the Student’s t-test in H (n = 6). **p < 0.01, *p < 0.05; ns, not significant. A ratio of 1.5-fold (>1.50 or <0.67) with a p-value < 0.05 was chosen as the cutoff for protein up- or downregulation in F (n = 3)