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. 2011 Dec 14;6(12):e29088. doi: 10.1371/journal.pone.0029088

Figure 1. Expression of LYN and -associated genes are androgen-regulated in LNCaP cells.

Figure 1

(A) Western analysis of LYN and actin (loading control) protein expression in cells grown in full medium, steroid-deplete medium, and following 24 hours of treatment with 10 nM R1881. (B) Ingenuity Pathway Analysis (IPA) of LYN-associated genes. Novel androgen-regulated genes identified in the present study are labelled in shades of red (up-regulated genes) or shades of green (down-regulated genes). (C) Expression of 12 novel androgen-regulated genes associated with LYN in steroid-deplete conditions (−, lanes 1–4) and after 24 hours of treatment with 10 nM R1881 (+, lanes 5–8) measured by multiplex RT-PCR. Levels of transcript expression were normalised to GAPDH levels, and data from experimental quadruplicates were used to obtain the mean ± SE for each transcript. A particularly high level of androgen-mediated gene activation was seen for the FZD5 gene, which encodes a transmembrane Wnt5a receptor. Other confirmed up-regulated genes included KITLG, which encodes the Kit ligand upstream of LYN in the pathway, and MTOR, which encodes a key regulator of cell growth. Similarly, we confirmed down-regulation of the novel androgen-regulated genes REDD1 (also known as DDIT4), which negatively regulates mTOR, and DEPDC6 which encodes an mTOR interacting protein.