Fig. 2: NAT10 controls nuclear acetyl-CoA production to activate the transcription of chemotherapy-resistant genes. | Cell Death & Disease

Fig. 2: NAT10 controls nuclear acetyl-CoA production to activate the transcription of chemotherapy-resistant genes.

From: Chemotherapy-induced acetylation of ACLY by NAT10 promotes its nuclear accumulation and acetyl-CoA production to drive chemoresistance in hepatocellular carcinoma

Fig. 2

A The enriched pathways of the NAT10 downregulated genes were plotted by KEGG enrichment analysis using the data obtained by RNA-seq of the NAT10 shRNA and control cells after doxorubicin treatment. B The mRNA levels of CYP2C9, CYP2C19, NEIL1, XRCC1, and PIK3R1 genes were evaluated by RT-qPCR in Huh7-NAT10 shRNA-1, Huh7-NAT10 shRNA-2 and Huh7-control shRNA cells after doxorubicin treatment. Data were analyzed by one-way ANOVA and presented as mean ± SEM (n = 3), ns denotes no significance, *P < 0.05, **P < 0.01. C Huh7 cells were transfected with Flag-vector, Flag-NAT10 or Flag-NAT10 G641E plasmids and treated with doxorubicin. The mRNA levels of CYP2C9, XRCC1 and PIK3R1 genes were evaluated by RT-qPCR. Data were analyzed by one-way ANOVA and presented as mean ± SEM (n = 3), ns denotes no significance, *P < 0.05. D Huh7-NAT10 shRNA-1, Huh7-NAT10 shRNA-2, and Huh7-control shRNA cells were treated with oxaliplatin and then were treated with actinomycin D (Act.D). These cells were collected at different time points for RNA extraction. The mRNA levels of PIK3R1 and CYP2C9 genes were detected by RT-qPCR. E, G The protein levels of PIK3R1, p-AKT, AKT, and CYP2C9 were detected by WB in Huh7-NAT10 shRNA-1, Huh7-NAT10 shRNA-2 and Huh7-control shRNA cells with oxaliplatin treatment. F, H Huh7 cells were transfected with Flag-vector, Flag-NAT10 or Flag-NAT10 G641E plasmids and treated with oxaliplatin. The protein levels of PIK3R1, p-AKT, AKT, and CYP2C9 were detected by WB. I The H3K27ac levels on the promoters of PIK3R1 and CYP2C9 genes were detected by ChIP in Huh7-NAT10 shRNA-1, Huh7-NAT10 shRNA-2, and Huh7-control shRNA cells with 10 μM oxaliplatin treatment. Data were analyzed by one-way ANOVA and presented as mean ± SEM (n = 3), ***P < 0.001. J The acetyl-CoA levels were analyzed in Huh7-NAT10 shRNA-1, Huh7-NAT10 shRNA-2, and Huh7-control shRNA cells with or without oxaliplatin treatment. Data were analyzed by one-way ANOVA and presented as mean ± SEM (n = 3), ***P < 0.001. K Cellular fraction was prepared in Huh7-NAT10 shRNA-1, Huh7-NAT10 shRNA-2, and Huh7-control shRNA cells with or without oxaliplatin treatment. L The acetyl-CoA levels were analyzed in K’s cellular fractions. Data were analyzed by one-way ANOVA and presented as mean ± SEM (n = 3), ns denotes no significance, ***P < 0.001.

Back to article page