Fig. 6

Proteomic analysis of differentially expressed proteins in colon cancer cells. a, The 4D-label-free quantitative proteomics by the gene ontology (GO) enrichment analysis. b, Subcellular localization of these differentially expressed proteins. c, The analysis identified statistics of differentially expressed proteins after PCSK9 knockdown in HCT116 cells. d, The volcano plot showed differential proteins associated with glycometabolism and immune reactions. e and f, Western blot. HCT116 and HT-29 cells were grown and transiently transfected with PCSK9 or control siRNA or with plasmids carrying PCSK9 cDNA or vector only and then subjected to Western blot analysis at 48 hs after transfaction. g, Western blot. HCT116 and HT-29 cells were grown and transiently transfected with PCSK9 or control siRNA or with plasmids carrying PCSK9 cDNA or vector only and then subjected to Western blot analysis of levels of protein lactylation. h, Concentration of lactate in the culture supernatant of HCT116 cells was established by Lactate Assay Kit. NC, negative control; KD, knockdown of PCSK9 cells transfected with PCSK9 siRNA; OE, overexpression of PCSK9 cells transfected with plasmids carrying PCSK9 cDNA. *p < 0.05; ***p < 0.001