Fig. 1: KAT8 regulates lipolysis accompanied by enhanced KAT8 acetylation after PA treatment in human CRC cells.

A HCT116 cells were treated with PA (0.2 mM) for 24 h, glucose starvation (GS) for 18 h, etoposide (40 μM) for 8 h, HU (2 mM) for 2 h and CPT (1 μM) for 1 h. Cells were lysed and subjected to quantify the amount of glycerol inside cells by glycerol colorimetric assay kit. B, E HCT116 cells were transfected with KAT8 siRNA (B) or KAT8 plasmid (E), and then treated with or without PA at 0.2 mM for 24 h. Oil red O staining was used to detect the accumulation of lipid droplets. C The siRNA efficiency of KAT8 was detected by real-time PCR. D, G HCT116 cells were treated as outlined above, cells were lysed and subjected to quantify the amount of glycerol inside cells by glycerol colorimetric assay kit. F The transfected efficiency of KAT8 was detected by Western blotting. H, I SW480 (H) or RKO (I) cells were treated as outlined above, cells were lysed and subjected to quantify the amount of glycerol inside cells by glycerol colorimetric assay kit. J, K HCT116 cells were treated with 0.2 mM PA for 0, 18 h and 24 h (J) or PA (0.1 mM and 0.2 mM) for 24 h (K). The mRNA expression of KAT8 was analyzed by real-time PCR. mRNA levels of the control sample were set as 1, and relative mRNA levels of the experimental samples were normalized to this control. L, M HCT116 cells were stimulated under the same condition above. The total protein expression of KAT8 was detected by Western blotting. β-actin was used as a loading control. N, O HCT116 (N), SW480 and RKO (O) cells were treated with or without 0.2 mM PA for 24 h, cell lysates were then extracted for a Co-IP assay to detect endogenous acetylation levels of KAT8 in these cells. The bar (−) represents the means (n = 3).