Fig. 4: Genetic knockout of cholesterol biosynthetic enzymes impairs self-renewal and tumorigenic potential of colon CSC-enriched spheroids. | Oncogene

Fig. 4: Genetic knockout of cholesterol biosynthetic enzymes impairs self-renewal and tumorigenic potential of colon CSC-enriched spheroids.

From: CRISPR screens identify cholesterol biosynthesis as a therapeutic target on stemness and drug resistance of colon cancer

Fig. 4

A Colon CSC-enriched spheroids were transduced with sgRNAs targeting HMGCR or FDPS, followed by puromycin selection for 6 days. Knockout efficacy was evaluated by western blot. B Cell viability, C cell apoptosis, and D sphere-formation assays of colon CSC-enriched spheroids transduced with sgRNAs targeting indicated genes as compared to control. E Representative images of POP66 spheres treated as indicated. F In vitro limiting dilution assays of colon CSC-enriched spheroids after depletion of HMGCR or FDPS. G Representative images of excised tumors from mice injected with POP92 or LS174T-S spheroids after HMGCR or FDPS knockout. Tumor weight and tumor volume were measured. H Rescue assay using cholesterol biosynthesis pathway metabolites (FPP, GGPP, cholesterol) in HMGCR-depleted CSC-enriched spheroids. I Representative images of POP92 spheres treated as indicated. Error bar, mean ± SD. ns not significant, *P < 0.05 (t-test for B, C, G, one-way ANOVA for H).

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