Fig 2: PI3K/Akt inhibitors confirmed that EEF1D could regulate PI3K/Akt signal pathway. A and B PI3K/Akt inhibitors would decrease viability of EEF1D knockout/knockdown cells less than control cells; A SKOV3/DDP cell lines, B SKOV3 cell lines. C and D PI3K/Akt inhibitors had the similar effects from knockout/knockdown cells; C SKOV3/DDP cell lines, D SKOV3 cell lines. Data are mean ± SD, the experiment was performed with sextuplicate in three independent sets, *P < 0.05, **P < 0.01 compared with DDP intervention alone in the same cell line; #P < 0.05, ##P < 0.01 compared with SKOV3 or SKOV3/DDP in the same drug intervention

Fig 3: How EEF1D regulates PI3K/OPTN/Akt signal pathway was explored by OPTN knockdown in SKOV3/DDP/KD cell line. Data are mean ± SD, the experiment was performed with sextuplicate in three independent sets, **P < 0.01 vs. scrambled shRNA or OPTN shRNA control, respectively; #P < 0.05 vs. scrambled shRNA in the same drug intervention

Fig 4: Time course of the phosphorylation of JAK2, STAT5, AKT, mTOR, MEK1/2, and ERK1/2 and the quantitative variation of PRLR and ER-a in Ishikawa cells after adding prolactin.(a) The maximal phosphorylation of JAK2 was observed in Ishikawa cells 1 hour after treatment with 100 ng/ml of prolactin.(b) The phosphorylation of STAT5 was not changed in Ishikawa cells after treatment with 100 ng/ml of prolactin.(c) The phosphorylation of AKT and mTOR was not changed in Ishikawa cells after treatment with 100 ng/ml of prolactin.(d) The maximal phosphorylation of MEK1/2 and ERK1/2 was observed in Ishikawa cells 5 minutes and 15 minutes after treatment with 100 ng/ml of prolactin, respectively.(e) The increased expression of PRLR of 110 kDa and ER-a was observed in Ishikawa cells 6 hours after treatment with 100 ng/ml of prolactin(f) The maximal phosphorylation of ERK1/2 was observed in Ishikawa cells 5 minutes and 15 minutes after treatment with 100 ng/ml of prolactin, and the phosphorylation of ERK1/2 was completely inhibited in Ishikawa cells pretreated with 10 µM of U0126 after treatment with 100 ng/ml of prolactin.(g) The increased expression of PRLR of 110 kDa and ER-a was observed in Ishikawa cells 6 hours after treatment with 100 ng/ml of prolactin, and the decreased expression of PRLR and ER-a was observed in Ishikawa cells pretreated with 10 µM of U0126 6 hours after treatment with 100 ng/ml of prolactin.

Fig 5: Reducing the expression of EEF1D gene regulated the contents and/or activities of proteins associated with PI3K/Akt signaling pathway, apoptosis and repair of DNA damage in xenografted tumor. A The related proteins in xenografted tumor tissues were detected with Western blotting, and ß-actin was used to show the similar amount of protein loaded in different lanes. B, D Relative intensities of protein bands in A were determined using Quantity-One software and normalized using ß-actin band intensity. C Akt phosphorylation levels were calculated according to the relative intensities of p-Akt/Akt bands, normalized using ß-actin. Data in B, C and D are presented as mean ± SD of six mice in each group. * P < 0.05, ** P < 0.01 vs. control; # P < 0.05, ## P < 0.01 vs. DDP group