Fig. 3

Multiple cancer-related pathways and receptors are activated in CerS4-overexpressing MCF-7 cells. (A) Well-known tumor-related pathway components (Akt/mTOR, NF-κB, and GSK3β/β-catenin) (left) and densitometric analysis (right) of the indicated protein in MCF-7 and MCF-7/CerS4 cells (n = 3) (B) β-catenin levels in nuclear and cytosol fractions were assessed in MCF-7 and CerS4-overexpressing MCF-7 cells (MCF-7/CerS4). (C) Relative ESR1, ESR2, and PGR mRNA expression levels in MCF-7 and MCF-7/CerS4 cells. (D) Representative western blots showing protein levels of ERα, PGR, and phosphorylated (Ser118, Ser167) ERα and (E) their densitometric analysis in MCF-7 and MCF-7/CerS4 cells (n = 3). (F) Increased expression of lipid-related genes in MCF-7/CerS4 cells compared with MCF-7 cells (n = 3). Two-tailed Student’s t-test was performed. Significance is indicated as follows: ^*P < 0.05, ^**P < 0.01, and ^***P < 0.001. Akt, protein kinase B; ERα, estrogen receptor alpha; ERK, extracellular signal-related kinase; FAS, fatty acid synthase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GSK3β, glycogen synthase kinase-3 betta; HA, hemi-agglutinin; HMGCR, 3-hydroxy-3-methylglutaryl-CoA reductase; LDLR, low-density lipoprotein receptor; mTOR, mammalian target of rapamycin; NF-κB, nuclear factor kappa B; PGR, progesterone receptor; S6K, ribosomal S6 kinase; SCD1, stearoyl-CoA desaturase 1; SREBP, sterol regulatory element–binding protein