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. 2017 Oct 20;18(11):904–915. doi: 10.1080/15384047.2017.1385678

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Figure 5. — HN1L regulates Ras/MAPK pathway via interacting with RASA4. (a) List of the top HN1L-interacting proteins from the public databases. Co-immunoprecipitation (b) and immunofluorescent double-staining (c) indicate the direct interaction of HN1L and RASA4 in A549 and HCC827 cells. (d) GST-Raf1-RBD fusion protein is used to bind the activated form of GTP-bound Ras, which can then be immunoprecipitated with glutathione resin. Ras activation levels in A549 and HCC827 cells with or without HN1L silencing are then determined in western blot using a Ras monoclonal antibody. (e) Immunofluorescent staining showed the reduction of membrane localization of Raf-1 after knockdown of HN1L. Scale bar, 10 μm.