Fig. 3

Increased succinylation leads to enhanced MGs’ inflammatory susceptibility, ROS over-production, and lipid metabolism disruption. (A) Schematic experimental workflow of Sirt5 KD BV2 cells with LPS treatment and further experiments. (B) Succinylation level between CTL and Sirt5 KD BV2 cells with LPS treatment in (a) total lysate and (b) mito lysate detected by WB. C. The glycolytic rate changes of CTL and Sirt5 KD cells following LPS treatment detected by the glycolytic rate test. D. The ATP production rate changes of CTL and Sirt5 KD cells following LPS treatment detected by the real-time ATP production rate assays. E. mtROS generation of CTL and Sirt5 KD cells treated with LPS detected by FC. F. Lipid accumulation of CTL and Sirt5 KD cells treated with LPS detected by FC. G. MDA concentration of CTL and Sirt5 KD cells treated with LPS detected by colorimetric method. H. Pro-inflammatory cytokines transcription levels of Il-1β, Il-6 and Tnf-α in the CTL and Sirt5 KD cells treated with LPS detected by qPCR. I. Schematic workflow of non-targeted metabolomics. J. OPLS-DA analysis of non-targeted metabolomic datasets between CTL and Sirt5 KD cells. (n = 6/group) K. Volcano plot showing different metabolites in CTL and Sirt5 KD cells (n = 6/group). L. The counts of up and downregulated of different metabolites classified by superfamily. M. KEGG enrichment of different metabolites in CTL and Sirt5 KD BV2 cells. Upregulated metabolites are in red, and downregulated genes are in blue. n = 3/group, * P < 0.05, ** P < 0.01, *** P < 0.005, **** P < 0.001