N-Acetyltransferase 1 Knockout Elevates Acetyl Coenzyme A Levels and Reduces Anchorage-Independent Growth in Human Breast Cancer Cell Lines

Figure 4

Doubling time and anchorage-dependent and anchorage-independent colony formation of parental and NAT1 KO clones for MDA-MB-231, MCF-7, and ZR-75-1 cell lines. (a) Doubling time was determined in MDA-MB-231, MCF-7, and ZR-75-1 parental (P) and gRNA #2 (2) and #5 (5) NAT1 KO clones. Each bar illustrates mean ± SEM for number of replicates (N). Doubling times differed significantly between parental and NAT1 KO cell lines for MDA-MB-231 (), and ZR-75-1 () cell lines. (b) Anchorage-dependent growth/colony formation was determined in MDA-MB-231, MCF-7, and ZR-75-1 parental (P) and NAT1 KO cell lines. Cells (300) were plated on plastic in triplicate and allowed to grow for 14 days before staining. Anchorage-dependent growth/colony formation between parental and NAT1 KO cells were not significantly different () for all cell lines, except for ZR-75-1. (c) Anchorage-independent/soft agar assays were completed in MDA-MB-231, MCF-7, and ZR-75-1 parental (P) and NAT1 KO cell lines. Cells (6000) plated in triplicate in soft agar were allowed to grow for 14 days before staining. The number of colonies formed in soft agar were significantly higher in parental MDA-MB-231 () MCF-7 () and ZR-75-1 () than their respective NAT1 KO cell lines.

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