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. 2021 Sep 9;3(3):zcab035. doi: 10.1093/narcan/zcab035

Figure 3.

Figure 3.

CPT1A is required for HR+ breast cancer cell line clonologenicity and mammosphere formation. (A) Silencing of CPT1A in MCF7 or (B) ZR751 dox-inducible shRNA expressing cells demonstrate reduced colony formation following dox treatment (2 μg/ml dox, 21 days); no change is seen in MCF7shNS or ZR751shNS cells +/-dox (C). Quantitation of colony formation assays (t-test) reported as the ratio of +/-dox. (D) Western blot analysis demonstrates relative Cpt1a overexpression in MCF10A cells following lentiviral transduction (MOI = 3, 72 h). (E) CPT1A overexpression in MCF10A cells result in increased colony formation capacity. (F) Quantitation of colony formation assay in (E). An unpaired-test was used for each analysis: ***p<0.0005; **p<0.005; mean and standard error are indicated. (G) MCF7 or (H) ZR751 cells expressing either sh1 or sh2 tet-inducible shRNA against CPT1A show reduced mammosphere formation following dox treatment (2 μg/ml; 21 days); no effect on mammosphere formation was apparent in parental or shNS expressing cells +/-dox. Quantification of mammosphere following shRNA-mediated silencing of CPT1A in (I) MCF7 or (J) ZR751 cells; data are presented relative to no dox treatment for each cell line. (K) CPT1A overexpression (MOI: 3, 10 days) induces mammosphere formation in MCF10A cells. (L) Quantification of (K) as relative mammosphere count. An unpaired t-test was performed for all assays.