Fig. 4

BME treatment inhibits phospholipids, iPLA2 activity and lipid raft. Cal27 and JHU022 cells were treated with BME for 30 h and lipid profile was analysed by electrospray ionization-tandem mass spectrometry (ESI-MS/MS). Representative mass- spectra showing a: phosphatidylcholine (PC), b: phosphatidylethanolamine (PE), and c: plasmenylethanolamine. d: Cal27 and JHU022 cells were treated with BME for 30 h and intracellular iPLA2 activity was assayed. Small bar indicates standard error (*, p < 0.05; **, p < 0.01; *** p < 0.001). e: Cal27 and JHU022 cells were treated with BME for 30 h and stained with antibody to Flotillin (red) and DAPI (blue). Representative confocal microscopic images showing reduced expression of Flotillin in BME treated cells compared to control cells. Magnifications 60X and scale bar 50 μm. f: Cell lysates from Cal27 and JHU022 with or without BME treatment for 30 h were subjected to Western blot analysis for Flotillin-1 using specific antibody. The membrane was reprobed with antibody to actin as an internal control. Quantitative of Western blot band intensities using Image-J software (shown in right). Small bar indicates standard error (**, p < 0.01; *** p < 0.001)