Fig. 6

eIF5A hypusination contributes to TCF4 protein translation. (A) TCF4 contains 7 conserved Pro-Pro motifs. (B) Expression of TCF4, AMD1, eIF5A and eIF5A hypusination was examined by Western blotting in BT549 cells with or without eIF5A knockout (eIF5A-KO) as well as eIF5A-KO-expressing BT549 cells with eIF5A-WT or eIF5A-K50R expression. (C) Expression of TCF4, AMD1, eIF5A and eIF5A hypusination was examined by Western blotting in BT549 cells with or without eIF5A knockout (eIF5A-KO) as well as eIF5A-KO-expressing BT549 cells with eIF5A-WT or eIF5A-K50R expression following treatment with or without spermidine (30ng/ml) for a period of 24 h. (D) Expression of AMD1, TCF4 and cyclinD mRNA was analyzed by quantitative real-time PCR in BT549 cells with stable empty vector or ADM1 expression as well as SUM159 cells with stable empty vector or knockdown of AMD1 expression. Data are shown as mean ± SD based on three independent experiments