Fig. 5
TPX2 expression is regulated by Hh-FOXM1 signaling. a. Protein expression levels of TPX2 and FOXM1 were assessed after stable knock down of FOXM1 in Huh7 and Hep3B cells using WB analysis. b. WB analysis to determine protein expression levels of TPX2 and FOXM1 in Huh7 and HepG2 cells stably over-expressing FOXM1. c-e. FOXM1 depletion suppresses tumor growth in vivo. Huh7 cell lines (2 × 107 cells) that expressing sh-FOXM1 or sh-Con were subcutaneously injected into nude mice on each side of the inguinal region. Mice were administered 2 μg/mL doxycycline and 5% sucrose in sterile drinking water. Xenografts were harvested after 3 weeks. Tumor sizes on either side were monitored every other day (c), and tumor size (d) and weight (e) are shown. Data are presented as mean ± SD (n = 5). *, p < 0.05; **, p < 0.01. f-g. FOXM1 depletion down-regulates TPX2 expression in vivo. Protein level of TPX2 in tumors were checked by western blot (f) and IHC (g). h. Huh7 cells transfected with shRNA-control or shRNA-FOXM1 were incubated with or without N-Shh conditional medium for 48 h and were harvested for WB analysis with the indicated antibodies. i. Huh7 cells stably infected with Lv-control or Lv-GLI2A were infected twice with shRNA-control or shRNA-FOXM1 and were harvested for WB with the indicated antibodies. j. Hep3B cells transfected with Lv-control or Lv-FOXM1 were treated with DMSO or 20 μM GANT61 for 48 h and were harvested for WB analysis with the indicated antibodies. Data was shown as mean ± SD (n = 3). *, p < 0.05; **, p < 0.01