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Figure 1


Fig. 1. Isolation of anti-CD36 antibodies blocking cytokine-induced macrophage fusion. (A) ThioM{Phi} were labelled with CFSE and PKH26 and fusion induced by exposure to IL-4 in the presence of supernatants from three individual hybridoma lines MF2, MF3 and MF4. Macrophage fusion is represented by co-localization of the red and green fluorescent labels (yellow). (B) Quantitation of co-localization in the presence of the purified anti-CD36 antibodies MF2, MF3 and MF4 (20 µg/ml). Means±s.d. of 21 measurements combined from three independent experiments. (C) MF2, MF3, MF4 or isotype control (IgG2a, IgG2b) antibodies were covalently coupled to protein G beads and used for immunoprecipitation from ThioM{Phi} lysates. Specific bands (~100 kDa) could be detected on the silver-stained gel for MF2, MF3 and MF4 but not for isotype control antibodies. (D) Transient transfection of CHO cells with mCD36-YFP or YFP control vector, stained with MF2, 3 and 4. Positive staining was detected via anti-rat Alexa 555 (red). Positive staining with MF2, 3 and 4 was only detected in CHO cells expressing the mCd36-YFP fusion protein. (E) Western Blot analysis of wild-type (WT) and CD36-KO macrophage lysates using MF3, MF2, MF4 and anti-β-actin antibodies. ***P<0.0001, Mann Whitney Test, two-tailed.





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