Fig. 4
From: Dual blockade of the lipid kinase PIP4Ks and mitotic pathways leads to cancer-selective lethality
Identification of positive cross-talk between Ras/Raf/MEK/ERK and PI3K/Akt/mTOR pathway via Ras˧PIK3IP1˧PI3K signaling network. a The PI3K network analysis using gene expression of PI3K regulators in normal (middle) and transformed BJ cells (right) treated with a131 at 5 μM for 24 h. PI3K regulators, including PIK3IP1, were identified to interact with PI3K with experimental support and high confidence from STRING. The per-sample expression profiles of PIK3IP1 are depicted in the heatmap (left). Color: negative log FDR (false discovery rate), coded from white to red in a scale from 0.15 to 5.37. Size: log ratio; Border: upregulation (red) or downregulation (blue); Shape: upstream (square), parallel (diamond) or downstream (circle). b, e qRT-PCR analysis of PIK3IP1 expression in BJ cell lines treated with a131 or a166 at 5 µM for 24 h. Mean values ± S.D. (n = 3). c Immunoblot analysis of normal BJ cells treated with 4-OHT [4HT(+)] for 24 h to activate H-RasV12-ER and subsequently with a131 (left) or transfected with indicated siRNAs for 48 h (right). Representative immunoblots shown (n = 3). d ChIP analysis of normal BJ cells treated with a131 using antibodies against RNA polymerase II (Pol II) for the PIK3IP1 gene promoter (n = 3). e Normal BJ cells treated with a131 were subsequently treated with MEK inhibitor U0126 (10 µM) for additional 2 h. Then, 4-OHT added to activate H-RasV12-ER for various time points. Where indicated, two-tailed unpaired t tests were performed to determine statistical significance
