Src specific activity in non-invasive (PZ-HVP-7, RWPE1), androgen-independent (CWR22Rv1), and invasive (DU145, PC3) cell lines.

<p>Src kinase specific activity was calculated by dividing Src activity (Fig. 2) by total Src protein content (Fig. 3). Src specific activity is significantly higher in aggressive than in non-cancer cell lines (P<0.0001). Error bars are SEM.</p

Src kinase activity in non-invasive (PZ-HPV-7, RWPE1), invasive (DU145, PC3), and androgen-independent (CWR22Rv1) cell lines.

<p>Grey bars are phosphorylation rates of peptide 1 by whole cell lysates (normalized by total protein extract amount). White bars are phosphorylation rates by cell lysates due to Src kinase alone after subtracting non-Src background phosphorylation of 1. Comparison between non-cancer (PZ-HPV-7, RPWE1) and aggressive cancer cell lines (CWR22Rv1, DU145, PC3) showed significant lowered levels of Src kinase activity associated with the later (p<0.001). All experiments were performed at least in triplicate. Error bars are SEM.</p

Src expression levels in non-invasive (PZ-HPV-7, RWPE1), androgen-independent (CWR22Rv1), and invasive (DU145, PC3) cell lines.

<p>(a) Prostate cell lysates were probed for total Src, pY416 Src and pY527 Src, where α-tubulin was used as the loading control. (b) Intensities of each band from the western blots were measured and normalized to the corresponding α-tubulin control, and then compared to cell line RWPE1 (RWPE1 as 1). Comparison between non-cancer (PZ-HPV-7, RPWE1) and aggressive cancer cell lines (CWR22Rv1, DU145, PC3) showed significant lowered levels of Src expression in the latter (p<0.001). Data are representative of at least three independent experiments and are mean with SEM.</p

Src-catalyzed phosphorylation rates, Src protein content, and Src phosphorylation status in the RWPE1-derived cell lines.

<p>Increasing invasive ability is plotted along the x-axis. (a) Grey bars are phosphorylation rates of peptide 1 by whole cell lysates (normalized by total protein content). White bars are phosphorylation rates by cell lysates due to Src kinase alone after subtracting non-Src background phosphorylation of 1. (b) Total Src content as determined by western blot analysis (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048867#pone.0048867.s005" target="_blank">Fig. S5</a>) (c) Src kinase specific activity as assessed by measured Src activity (Fig. 6A) divided total Src protein content (Fig. 6B). (d) pY416 levels were derived from the band intensities in the western blots (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048867#pone.0048867.s004" target="_blank">Fig. S4</a>), normalized to the corresponding α-tubulin control, and then compared to cell line RWPE1 (RWPE1 as 1). Data are representative of at least three independent experiments and are shown as mean with SEM.</p

Assessment of Src kinase catalytic activity.

<p>(a) CE-LIF separation and visualization of the Src peptide substrate <b>1</b> and its chemically synthesized phosphorylated counterpart <b>2</b>. (b) Src kinase-catalyzed phosphorylation of peptide <b>1</b> as a function of time as assessed by CE-LIF.</p

DataSheet1_Expression profiling of ALOG family genes during inflorescence development and abiotic stress responses in rice (Oryza sativa L.).ZIP

The ALOG (Arabidopsis LSH1 and Oryza G1) family proteins, namely, DUF640 domain-containing proteins, have been reported to function as transcription factors in various plants. However, the understanding of the response and function of ALOG family genes during reproductive development and under abiotic stress is still largely limited. In this study, we comprehensively analyzed the structural characteristics of ALOG family proteins and their expression profiles during inflorescence development and under abiotic stress in rice. The results showed that OsG1/OsG1L1/2/3/4/5/6/7/8/9 all had four conserved helical structures and an inserted Zinc-Ribbon (ZnR), the other four proteins OsG1L10/11/12/13 lacked complete Helix-1 and Helix-2. In the ALOG gene promoters, there were abundant cis-acting elements, including ABA, MeJA, and drought-responsive elements. Most ALOG genes show a decrease in expression levels within 24 h under ABA and drought treatments, while OsG1L2 expression levels show an upregulated trend under ABA and drought treatments. The expression analysis at different stages of inflorescence development indicated that OsG1L1/2/3/8/11 were mainly expressed in the P1 stage; in the P4 stage, OsG1/OsG1L4/5/9/12 had a higher expression level. These results lay a good foundation for further studying the expression of rice ALOG family genes under abiotic stresses, and provide important experimental support for their functional research.</p