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. 2017 Nov 26;8(65):108292–108302. doi: 10.18632/oncotarget.22660

Figure 4. SLPI physically interacts with Rb.

Figure 4

A., B. Co-immunoprecipitation of SLPI and Rb. Total cell lysates from two different cell lines (MDA-MB-231 and HEK293T) were immunoprecipitated and analyzed by immunoblotting with antibodies against SLPI and Rb. Irrelevant isotype-matched IgG antibody was use as a control. C. Interaction between SLPI and Rb assessed by mammalian two-hybrid system. We cloned Rb into pBIND, and SLPI and FoxM1 into pACT vectors to generate fusion proteins. Forty-eight hours after transfection the cells were lysed and the amount of luciferases were quantitated using the Dual-Luciferase Reporter Assay System. pACT-FoxM1 and pBIND-Rb were used as a positive control for established physical interaction. Error bars represent SEM; n = 3, * P < 0.01. D. Localization of SLPI protein in breast cancer cells. The images were taken using an Olympus U-TBI90 microscope with 60x Objective. The images show immunostaining with anti-SLPI antibodies (red) and DAPI (blue).