Figure 6.
Effect of IFN-γ on OCTN2 level in cell and exosomes. HEK293 cells were treated with 50ng/ml IFN-γ for 48 hours. After incubation, cells were harvested. Exosomes were isolated from the medium of the same culture. (a) Both cell (upper panel) and exosomal (lower panel) protein extracts deriving from IFN-γ treated (right line) or untreated (left line) culture was analyzed by WB using the antibody against OCTN2. The immunostaining of cell extract with the antibody against actin was used as loading control. Exosomal OCTN2 was normalized using TSG101 by loading on SDS-PAGE an equal amount of the same exosomal extracts used to test the OCTN2 expression. Cell lysate and exosomal extract were subjected to WB. The membrane was cut for incubation with the different antibodies (b) Transport was started adding 50 µM 3H-carnitine to proteoliposomes and stopped after 60 min. Proteoliposome was reconstituted using either exosomal (gray bar) or cell (white bar) protein extracts. The values are the mean ± S.D. from three independent experiments. (*) Significantly different as estimated by the Student’s t test (p < 0.05).