Fig. 1

The crotonylome in HCC tissues and cells. A, B Immunohistochemistry (IHC) assays showing that pan-crotonylation was higher in HCC patients with metastasis. The data are presented as the means ± SD. (Student’s t test). C Patients with high pan-crotonylation levels had a shorter disease-free survival period than those with low pan-crotonylation levels. D, E IHC assays of 100 HCC tissues showing that pan-crotonylation was correlated with E-Cadherin and N-Cadherin. F Flowchart showing the experimental procedure followed for identifying crotonylated proteins through stable isotope labeling by amino acids in cell culture (SILAC) and liquid chromatography with tandem mass spectrometry (LC–MS/MS). G Subcellular localization of the identified crotonylated proteins. H Gene set enrichment analysis (GSEA) showing the five most enriched pathways (p < 0.05). I The difference in protein expression and crotonylation between MHCC-97H and MHCC-97L cells. The level of protein expression and degree of crotonylation were increased in MHCC-97H cells (-lg false discovery rate [FDR] > 0). J Scatter diagram showing the differentially crotonylated sites in the MHCC-97H and MHCC-97L cell lines (p < 0.05). K Western blotting (WB) verified that the total crotonylation level was higher in the MHCC-97H cell line