Figure 1.
Transient CHKA silencing efficiently downmodulates ChoK-alpha and intracellular PCho content in EOC cell lines. (A): CHKA silencing was confirmed at mRNA (RT-qPCR, upper panel; data are the mean±s.d. of five independent experiments) and protein expression level (lower panel: western blotting of a representative experiment is reported). (B): RT-qPCR for ChoK β-isoform. Data are the mean±s.d. of five independent experiments. GAPDH was amplified as endogenous control and I64-hTERT cells preparations were used as calibrator. (C): Representative examples of alterations induced by siCHKA on the 1H-MRS tCho profile and its PCho component. Black lines: control cells; grey lines: silenced cells. Inset in lower panel: western blot of control and siCHKA EOC cells probed with specific antibody to ChoK-alpha. GPC, glycerophosphocholine; PCho, phosphocholine; tCho, total choline-containing compounds. The PCho/tCho ratio was 0.75±0.20 in both cell types before silencing.