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. 2015 Dec 22;7(4):4632–4646. doi: 10.18632/oncotarget.6728

Figure 6. RKIP directly interacts with full-length Notch1.

Figure 6

(A) The left panel indicates the input of RKIP and Notch1 in FLAG-tagged, RKIP-overexpressing cells. Total cell lysates (1 mg) of FLAG-tagged, RKIP-overexpressing H1299 or HeLa cells and control cells expressing pcDNA were immunoprecipitated (IP) using anti-FLAG antibodies (M2 beads), and the bound Notch1 proteins were detected by immunoblotting (IB) with anti-Notch1 antibodies. (B) Co-localization of RKIP and Notch1 using immunohistochemistry. FLAG-tagged, RKIP-overexpressing H1299 cells were fixed, stained with both FITC-conjugated anti-RKIP antibodies and Cy5-conjugated anti-Notch1 antibodies, and visualized under the fluorescent microscope. The nuclei were stained with H33342 dye. White dotted boxed regions correspond to the higher magnification images below. (C) Co-localization of EGFP-RKIP and Notch1. EGFP-tagged, RKIP-overexpressing H1299 cells were fixed, stained with both GFP and Cy5-conjugated anti-Notch1 antibodies, and visualized under the fluorescent microscope.