Fig. 2

Localization of LIP in the lipid raft microdomains of cancer cell membranes. (a) A total of 5 × 10⁴ cancer cells or normal cells were incubated with Alexa488-tagged LIP (1 μg/mL) at 37 °C for 30 min and then subjected to flow cytometric analysis. The upper panels and lower panels show the results before and after LIP treatment, respectively. (b) The cells were observed and photographed using a Zeiss LSM 780 inverted microscope (magnification: 63×). (c) The cells were incubated with LIP (1 μg/mL) at 37 °C for 30 min. The cell membranes and culture medium were independently collected, resolved by SDS-PAGE and probed by western blotting using anti-LIP antibodies. (d) MCF-7, HepG2, H293T and MCF-10A cells were stained with Alexa555-cholera toxin subunit B (CT-B) prior to staining with Alexa488-tagged LIP. The CT-B was used following the instructions from Thermo Fisher Scientific. The cells were observed and photographed by 3D-SIM superresolution microscopy