Fig. 1

ACLY is upregulated by SEC63 in response to ER stress in HCC cells. A Venn diagrams of overlapped differentially expressed Acetyl-CoA-Related Genes (ARGs) among TCGA-LIHC, ICGC-LIRI-JP and GSE101728 cohorts. B Huh7 cells were treated with tunicamycin (TM, 5 μg/mL), or thapsigarin (TG, 1 μM) for 4 h. Then, the indicated proteins were detected by western blot. C Representative IHC staining of ACLY in HCC and adjacent non-tumorous tissues. Summary of ACLY expression in 139 cases of HCC and 20 cases of non-tumorous counterparts. Scale bars, 50 μm. D The overall survival (OS) or disease-free survival (DFS) of patients with HCC exhibiting different levels of ACLY staining was determined by Kaplan–Meier analysis. E The survival curves of the HCC patients with different ACLY mRNA levels from TCGA-LIHC cohort were calculated. F Huh7 cells were treated with TM (5 μg/mL) or TG (1 μM) for the indicated time points. Then, the expression of ACLY was evaluated by western blot. G The ER-related proteins among ACLY-interacting partners were identified by Co-IP-mass spectrometry. H Huh7 cell lysates were immunoprecipitated with anti-SEC63 or anti-ACLY antibody. Immuno-precipitates were immunoblotted as indicated. I Huh7 cells were treated with TM (5 μg/mL) or TG (1 μM) for 8 h and immunoprecipitation was performed with anti-SEC63 antibody. J, K Huh7 cells were transfected with ACLY or SEC63 deletion mutants and treated with MG132. Co-IP was performed using the anti-Flag antibodies. L The control and SEC63-depleted cells were treated with TM (5 μg/mL) or TG (1 μM) for the indicated time points, and the expression of ACLY and SEC63 was detected by immunoblotting