Fig. 2

Establishment of a CerS4 overexpressing MCF-7 cell line. MCF-7 cells overexpressing CerS4 were selected by G418 treatment. (A) Relative mRNA expression levels of various CerS family members in MCF-7 and CerS4-overexpressing MCF-7 cells (MCF-7/CerS4) (n = 3). (B) Ceramide levels in MCF-7 and CerS4-overexpressing MCF-7 cells (MCF-7/CerS4) using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) (n = 3). (C) Representative western blots showing protein expression levels of various CerS family members (left) and densitometric analysis (right) of CerS protein levels in MCF-7 and MCF-7/CerS4 cells (n = 3). (D) Cell proliferation in MCF-7 and MCF-7/CerS4 cells (n = 3). A two-tailed Student’s t-test was performed for (A, B, and C). A two-way ANOVA and a Tukey post hoc test was performed for (D). Significance is indicated as follows: ^*P < 0.05, ^**P < 0.01, and ^***P < 0.001