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. 2023 Dec 14;25(1):378–403. doi: 10.1038/s44319-023-00012-6

Figure EV5. NME4 regulates coenzyme A metabolism by interacting with the key enzymes in the pathway.

Figure EV5

(A) HEK293T cells were cotransfected with Myc-tagged NME4 and C-terminal SFB (SFB)-tagged candidate genes, as indicated. The cell lysates were incubated with immunoglobulin G (IgG) control and antibodies against Flag. (B) HEK293T cell lysates were incubated with immunoglobulin G (IgG) control and antibodies against NME4. (C,D) Wild-type and NME4 KO Bel-7402 cells were treated with PO for 24 h. The acetyl-CoA (C) and malonyl-CoA (D) levels were determined by targeted metabolite analysis. Biological replicates, n = 3. (E) Wild-type and NME4 KO Bel-7402 cells were treated with PO for 24 h. ATP levels was measured by kit. Biological replicates, n = 3. (F) Wild-type and NME4-overexpression SK-Hep1 cells were treated with PO for 24 h. ATP levels was measured by kit. Biological replicates, n = 3. (G) Relative mRNA levels of SREBP1C and CHREBP were measured by RT‒qPCR. Biological replicates, n = 3. (H) Relative mRNA levels of key genes involved in de novo lipogenesis, triglyceride synthesis and triglyceride breakdown were measured by RT‒QPCR in wild-type and NME4 KO Bel-7402 cells. Biological replicates, n = 3. Data information: (C–H) data are presented as mean ± SEM. *P values ≤  0.05, **P values ≤  0.01, ***P values ≤  0.001, NS -P values >  0.05 (Student’s t test, unpaired).