Effect of crizotinib and alectinib on stimulus preference of retinal ganglion cells.

<p>(A and B) Post-stimulus time histogram before (pre), during (crizotinib), and after (wash) the application of 1.0μM of crizotinib in an OFF-cell (A) and in an ON-OFF cell (B). The bin size for the histogram was 50 ms. (C and D) Cumulative distributions of the differences in <i>SPI</i> for crizotinib and alectinib. The difference in <i>SPI</i> was calculated from the <i>SPI</i> values evaluated before and during drug application. The cells were divided into “Decrease” type (C) and “Increase” type (D) according to the change in the <i>SPI</i>. The difference in (D) was an absolute value. (E and F) STA before (pre), during (crizotinib), and after (wash) the application of crizotinib in the cells shown in Figs 2A (E) and B (F). The amplitude “A” was defined as the difference in light intensity between the maximum and the minimum (double-headed arrow). (G and H) Plot of the amplitude “A” before (pre) and during drug application (drug) for the cells shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0135521#pone.0135521.t004" target="_blank">Table 4</a>. The cells were divided into “Decrease” type (G) and “Increase” type (H). **<i>P</i> < 0.01; *** <i>P</i> < 0.001; paired <i>t</i>-test.</p

Effects of crizotinib or alectinib on firing rate.

<p>Effects of crizotinib or alectinib on firing rate.</p

Effects of crizotinib or alectinib on <i>SPI</i>.

<p>Effects of crizotinib or alectinib on <i>SPI</i>.</p

Effect of crizotinib (A-C) or alectinib (D-F) on firing rate of retinal ganglion cells.

<p>The timing of the drug application is indicated by the bar above the traces. The drug concentration was 1.0μM for both crizotinib and alectinib. The results for no change-type (A, D), increase-type (B, E), and decrease-type (C, F) cells are shown. The retina was repeatedly exposed to a set of light stimuli (1-s bright stimulation and 1-s dark stimulation at a frequency of 0.5 Hz). The ordinate shows the average firing rate for 10 cycles of stimuli (20 s).</p

Oligonucleotide primers used for qPCR.

<p>Oligonucleotide primers used for qPCR.</p

L'Écho des imprimeurs et des libraires

01 novembre 18421842/11/01 (A5)-1842/11/01.Note : Prospectus

Combinatory effects with ZOL and CDDP in an orthotopic animal model.

<p>MSTO-211H cells (1×10⁶) were inoculated into the pleural cavity of BALB/c <i>nu/nu</i> mice (n = 6) (day 1), and then ZOL (25 µg, day 3) was administrated into the pleural cavity and/or CDDP (100 µg, day 5) into the peritoneal cavity (CDDP). PBS was used as a control. Tumor weights were measured on day 24. The SE bars are also shown. * P<0.05, ** P<0.01.</p

ZOL-induced up-regulation of p53 and knockdown of the p53 expressions with siRNA.

<p>(A, B) CDDP-treated (20 µM) and ZOL-treated (48 h) cells were subjected to Western blot analysis and probed with antibodies as indicated. Actin was used as a loading control. (C) Cells were treated with CDDP and/or ZOL for 48 h at the indicated concentrations and the expression levels of phosphorylated p53 were examined. (D) Cells were transfected with p53-targeted siRNA (p53-siRNA) or non-targeted control siRNA (Control) for 24 h and then treated with ZOL (50 µM) for 48 h. The lysate was subjected to Western blot analysis. (E) Cells were transfected with siRNA as indicted and were treated with ZOL for 3 days. The cell viabilities were measured with the WST assay and means of triplicated samples with the SD bars are shown. (F) Flow cytometrical analyses of MSTO-211H cells that were transfected with respective siRNA for 24 h and then treated with ZOL (50 µM) for 48 h. (G, H) Cells transfected with p53-siRNA were treated with different doses of ZOL and CDDP as indicated for 3 days and the CI values based on the cell viabilities were calculated at different Fa points with CalcuSyn software.</p

Cell cycle distribution of ZOL-treated cells.

<p>MSTO-211H cells were treated with or without ZOL (at 10 or 50 µM) for 24–72 h. Cell cycle was analyzed with flow cytometry.</p

Interferon-β Produces Synergistic Combinatory Anti-Tumor Effects with Cisplatin or Pemetrexed on Mesothelioma Cells

<div><p>Interferons (IFNs) have been tested for the therapeutic effects in various types of malignancy, but mechanisms of the anti-tumors effects and the differential biological activities among IFN members are dependent on respective cell types. In this study, we examined growth inhibitory activities of type I and III IFNs on 5 kinds of human mesothelioma cells bearing wild-type <i>p53</i> gene, and showed that type I IFNs but not type III IFNs decreased the cell viabilities. Moreover, growth inhibitory activities and up-regulated expression levels of the major histocompatibility complexes class I antigens were greater with IFN-β than with IFN-α treatments. Cell cycle analyses demonstrated that type I IFNs increased S- and G2/M-phase populations, and subsequently sub-G1-phase fractions. The cell cycle changes were also greater with IFN-β than IFN-α treatments, and these data collectively showed that IFN-β had stronger biological activities than IFN-α in mesothelioma. Type I IFNs-treated cells increased p53 expression and the phosphorylation levels, and activated apoptotic pathways. A combinatory use of IFN-β and cisplatin or pemetrexed, both of which are the current first-line chemotherapeutic agents for mesothelioma, produced synergistic anti-tumor effects, which were also evidenced by increased sub-G1-phase fractions. These data demonstrated firstly to our knowledge that IFN-β produced synergistic anti-tumor effects with cisplatin or pemetrexed on mesothelioma through up-regulated p53 expression.</p> </div