Fig 2: Study of the phosphatidylcholine biosynthetic pathway in LN-FLU and PC3 prostate cells. (A) Levels of CTP:choline cytidylyltransferase (CCTα) determined via Western blot and levels of CCTα mRNA determined via qPCR. Densitometric analysis of the Western blot bands is shown on the right of the image. Results are shown as the mean ± SD of three independent experiments. (B) Labeling of CCTα (green) in LNCaP, LN-FLU, and PC3 cells observed with fluorescence microscopy. Nuclei were labeled with DAPI (blue). A representative image of two different experiments is shown. (C) Levels of choline kinase (ChoK) determined via Western blot and levels of ChoK mRNA determined via qPCR. Densitometric analysis of the Western blot bands is shown on the right of the image. (D) Quantification of total phosphatidylcholine in LNCaP and LN-FLU cells using a phosphatidylcholine fluorometric assay. Phosphatidylcholine nanomoles were normalized by the amount of protein. (E) Diagram of phosphatidylcholine biosynthesis showing the metabolites and enzymes that we found down (red arrows) in LN-FLU cells compared to LNCaP cells. * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 indicate significant differences between LNCaP and LN-FLU cells via two-way ANOVA and Sidak’s multiple comparisons test.