Fig. 2

ACLY deficiency weakened lipid formation and metastasis of colorectal cancer cell. a Western blot analysis to access the efficiency of ACLY knockout in HCT116 cells and ACLY knockdown in RKO cells. Actin was the loading control. Ctrl: the normal control, KO: ACLY-knockout. #1-#3: the signal-cell clones that did not knock out ACLY in HCT116 cells. shNC: RKO signal-cell clones infected with negative control shRNA virus. shACLY: RKO signal-cell clones infected with shRNA-ACLY virus. b Real-time PCR analysis to qualify the changes of mRNA expression of the major lipid synthase when ACLY deficiency in HCT116 and RKO cells. SCD1: stearoyl-CoA desaturase 1, FASN: fatty acid synthase, HMGCR: 3-hydroxy-3-methylglutaryl-CoA reductase. c, d The quantitative detection of triglyceride and cholesterol when ACLY deficiency in HCT116 and RKO cells. e HCT116 cells stably knocked out ACLY and the control cells were used for Oil Red O staining (magnification, 200×). f, j Wound healing assays to access the effects of ACLY deficiency on the migration ability of HCT116 cells. g, h Transwell invasion assays to evaluate the effects of ACLY deficiency on the invasion ability of HCT116 and RKO cells. The quantitative analysis of cells across the transwell membrane (h and i). The quantitative analysis of wound healing rates (j)