Fig. 7
G6PD is critical for Plk1-mediated tumor proliferation in vivo. a, b HeLa cells stably expressing shG6PD were infected with viruses expressing pSin-3xFlag-G6PD wild type or mutants together with viruses expressing NTC (a) or shPlk1 (b) as indicated. Cell numbers were counted by trypan blue staining. n = 3 biologically independent replicates. Data were presented as mean ± s.d. *P < 0.05 as compared between indicated groups by two-sided Student’s t-test. c HeLa cells stably expressing shG6PD were infected with viruses expressing pSin-3xFlag-G6PD wild type and mutants together with viruses expressing tet-inducible shPlk1 as indicated. Cells were treated with 0.1 μg/ml doxycycline and cell numbers were counted by trypan blue staining. n = 3 biologically independent replicates. Data were presented as mean ± s.d. *P < 0.05 as compared between indicated groups by two-sided Student’s t-test. d–g HeLa cells stably expressing shG6PD were infected with viruses expressing pSin-3xFlag-G6PD wild type and mutants together with viruses expressing tet-inducible shPlk1 as indicated. Equal numbers of cells were subcutaneously injected into nude mice (n = 5 for each group). The mice were treated with or without doxycycline (1 mg/ml) in drinking water staring from 1 day before inoculation. The doxycycline-containing water was freshly prepared every other day. Tumor growth was measured starting from 10 days after inoculation (d, e). Tumors were extracted and compared at the end of the experiment (f, g). n = 5 for each group. Data were presented as mean ± s.d. *P < 0.05 as compared between indicated groups by two-sided Student’s t-test. h Protein levels of Plk1 and G6PD were determined by western blot using the lysates of tumor tissues from each group as in f using anti-Plk1 and anti-Flag. β-actin served as loading control
