RT-qPCR analysis of mRNA expression in iPS cells with or without maxadilan.

<p>Comparison of relative gene expression levels of Nanog, OCT4, PAX6, NESTIN, UTF1, SOX2, Rex1 and TERT in iPS cells between the control group and iPS cells treated with 100 nM maxadilan by RT-qPCR analysis. iPS cells that were not treated with maxadilan served as the control. Values are expressed as the mean of relative gene expression levels ± S.E.M of three independent experiments. There are no significant differences between the two groups (<i>p</i>>0.05 with the Student's <i>t</i>-test).</p

RT-qPCR analysis of mRNA expression in EBs from iPS cells with or without maxadilan.

<p>RT-qPCR analysis comparing the relative gene expression levels of PAX6 and NESTIN in the EBs from the control group and from iPS cells treated with 100 nM maxadilan. iPS cells that were not pretreated with maxadilan served as the control. Values are expressed as the mean of relative gene expression levels ± S.E.M of three independent experiments. There are no significant differences between the two groups (<i>p</i>>0.05 with the Student's <i>t</i>-test).</p

PAC1 mRNA and protein were expressed in iPS cells.

<p>The expression of PAC1 mRNA in iPS cells by RT-PCR analysis (a). The expression of PAC1 protein in iPS cells by western blot analysis showed that there are three kinds of PAC1 isoforms (b). The molecular weights of these PAC1 isoforms (1, 2 and 3) vary from about 50 kDa to 80 kDa.</p

List of primers.

<p>List of primers.</p

Effects of maxadilan at various concentrations in iPS cells exposed to UVC.

<p>Cell viability in iPS cells that were exposed to 50 J/m², 75 J/m² and 100 J/m² UVC and treated with maxadilan (at 0 nM, 3 nM, 5 nM, 10 nM, 20 nM, 30 nM and 50 nM) was detected by WST-8 analysis. iPS cells without UVC irradiation were used as control. Values are expressed as the mean of OD ± SEM of three independent experiments. ** <i>P</i><0.01 <i>vs.</i> control group using Dunnett's test. #<i>P</i><0.05 or ##<i>P</i><0.01 <i>vs.</i> UVC+0 nM Maxadilan group using Dunnett's test.</p

Comparison of protein fluorescence of Nanog and OCT4 in iPS cells with or without maxadilan.

<p>Comparison of fluorescence OD values of Nanog and OCT4 in iPS cells of the control group and in iPS cells treated with 100 nM maxadilan by microplate reader analysis. iPS cells that were not treated with maxadilan served as the control. Values are expressed as the mean of OD ± SEM of three independent experiments. There are no significant differences between the two groups (<i>p</i>>0.05 with the Student's <i>t</i>-test).</p

RT-PCR analysis of mRNA expression in iPS cells with or without maxadilan.

<p>RT-PCR analysis of Nanog, OCT4, PAX6, NESTIN, UTF1, SOX2, Rex1, and TERT mRNA expression in iPS cells of both the control group (a) and iPS cells treated with 100 nM maxadilan (b). iPS cells that were not treated with maxadilan served as the control.</p

Comparison of protein expression levels in iPS cells with or without maxadilan.

<p>Comparison of protein expression levels of Nanog, OCT4 and SOX2 in iPS cells between the control group and iPS cells treated with 100 nM maxadilan by western blot analysis. iPS cells that were not treated with maxadilan served as the control. Results are normalized to β-actin. Values are expressed as the mean ± S.E.M of three independent experiments. There are no significant differences between the two groups (p>0.05 with the Student's t-test).</p

Assessment of caspase-3 and caspase-9 activities in iPS cells with various treatments.

<p>Caspase-3 activity (A) and caspase-9 activity (B) in iPS cells that were exposed to 100 J/m² UVC and treated with maxadilan (at 0 nM and 30 nM) were measured by caspase-3 and caspase-9 treatment. iPS cells that did not undergo UVC irradiation were used as control. The results are expressed as the mean of OD ± S.E.M of three independent experiments.*<i>P</i><0.05 or **<i>P</i><0.01 <i>vs.</i> control group using Dunnett's test. #<i>P</i><0.05 <i>vs.</i> UVC+0 nM maxadilan group using the Student's <i>t</i>-test.</p

Comparison of Fluorescein-dUTP in iPS cells with various treatments by TUNEL assay.

<p>Fluorescein-dUTP of iPS cells that were exposed to 100 J/m² UVC and treated with maxadilan (at 0 nM and 30 nM) was detected by TUNEL assays. iPS cells that did not receive UVC radiation were used as control. Fluorescence values are given in arbitrary units and are expressed as the mean ± S.E.M of three independent experiments. **<i>P</i><0.01 <i>vs.</i> control group using Dunnett's test. #<i>P</i><0.05 Vs UVC+0 nM maxadilan group with the Student's <i>t</i>-test.</p