Mitochondrial FAO inhibition activates the mTORC1 pathway dependent on the acetyltransferase Gcn5.A, the effects of mitochondrial FAO inhibition on mRNA expression levels of acetyltransferase family related genes in ZFL cells. As shown in the heat map. N = 5. B, the effects of mitochondrial FAO inhibition on the protein expression levels of key acetyltransferases P300, Gcn5, and Pcaf in ZFL cells. N = 3. C, the effect of Gcn5 inhibitor (CPTH6, 1 μM) treatment on mTORC1 activity (p-S6kThr389 and p-S6Ser235/236) caused by mitochondrial FAO inhibition. The CN- (normal medium), MD- (normal medium containing 1 mM MD), and MD + CPTH6- (normal medium containing 1 mM MD and 1 μM CPTH6) treated cells were lysed and Western blotted after 48 h. N = 3. D, the 3D spatial structures of zebrafish Gcn5 and Raptor were obtained from SWISS-MODE and the predicted binding complex model of Raptor–Gcn5 by using the PDBePISA online protein docking tool. Note: different atoms are marked with different colors, gray is hydrogen, green is carbon, red is oxygen, and blue is nitrogen. E, the interface area and free energy of the predicted complex was analyzed and shown in the table. The larger the interface area implies the easier the proteins bind to each other. Negative free energy indicates that the protein can bind stably. F, immunoprecipitation reflects the protein interactions between Raptor and Gcn5 in ZFL cells. Data represent mean ± SD. ∗p < 0.05 and ∗∗p < 0.01. CN, control; FAO, fatty acid oxidation; Gcn5, general control nondepressible 5; MD, mildronate; mTORC1, mechanistic target of rapamycin complex 1; Pcaf, P300/Creb-binding protein–associated factor; ZFL, zebrafish liver.
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