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. 2016 Mar 2;7(16):21786–21798. doi: 10.18632/oncotarget.7846

Figure 6. Pin1 silencing impairs tumor growth and dissemination.

Figure 6

A. RT-qPCR of Pin1 mRNA. B. Immunoblot analysis of Pin1 protein levels, respectively, in a primary Eμ-myc lymphoma infected with conditional shRNAs targeting either Pin1 (shPin1) or renilla luciferase (shRen). The shRNAs were induced by supplementing cells with 1 μg/mL doxycycline for 24 hours. The mRNA data represent the averages ±s.d. of three technical replicates, all normalized to the housekeeper TBP and to the mock-treated shRen lymphoma. Vinculin was used as a loading control. C. Growth of two independent primary lymphoma populations (Ly27 and Ly28) infected with either shRen or shPin1, and cultured with (dox) or without (mock) doxycycline. D., E. Tumor-free survival in mice transplanted with an shRen or shPin1-bearing lymphoma (Ly28). Doxycycline was administered either continuously from the time of transplantation (D.) or following detection of tumor masses, 18 days after transplantation (E.). F. Dot plot showing the residual percentage of GFP-positive B220+ tumor cells detected in the tumor-infiltrated lymph nodes of animals transplanted with shPin1 or shRen lymphomas. Red bars indicate the average values. As in D., E., Mice were fed with doxycycline starting from days 0 or 18, as indicated. GFP serves as a marker for the doxycycline-dependent induction of the shRNAs. * p=0.014; ** p=0.0022; *** p=0.0026 (Log-rank, mantle-cox); # p=0.002, ## p=0.003 (t-test).