br Experimental design materials and methods Kinetic Fitting

Experimental design, materials and methods Kinetic Fitting: we used a fitting procedure that minimizes the error between the experimental data (concentration of the final double stranded complex as a function of time) and the solution of the model as obtained with an ordinary differential equation (ODE) integrator (Odeint from the Scipy library) [2]. UV-Thermal Melting Curves: were performed in 10mM cacodylate buffer (pH 7.2) and 100mM mixture of LiCl+KCl for quadruplex systems and 100mM LiCl for duplex structures. DNA systems were prepared at 4μM in the corresponding buffer and then denatured at 90°C for 5min and cooled down rapidly in ice. Thermal denaturation curves were obtained with a SAFAS spectrophotometer using quartz optical cells of 1cm pathlength using a scan rate of 0.3°C per min and following the variation of UV mtor pathway at 295nm and 260nm. CD experiments were performed with a JASCO J-815 spectropolarimeter equipped with a JASCO CDF-426S Peltier temperature controller, using quartz cells of 1cm path length. The scans were recorded from 210 to 335nm wavelength with the following parameters: 0.5nm data pitch, 2nm bandwidth, 100nmmin−1 scanning speed, and are the result of 3 accumulations.
Acknowledgments O.M. is the recipient of a post-doctoral fellowship from the fondation ARC. This work was supported by the Aquitaine regional council, PIA Nanobiotech program (Vibbnano #ANR-10-NANO-04) and ANR grants Quarpdiem [ANR-12-BSV8–0008–01] and Oligoswitch [ANR-12-IS07–0001].
Value of the data
Data The present DiB paper shows immunohistochemistry of redoxins including peroxiredoxins (Prdx1–6), glutaredoxins (Glrx1, 2, 3, 5), thioredoxins (Txn1, 2) and thioredoxin reductases (Txnrd1, 2) in the DRGs Figs. 1 and 2), spinal cord (Figs. 1, 2 and 7), sciatic nerve (Figs. 3–5, quantification Fig. 6) and thalamus (Fig. 8) in naïve mice and 7 days after Spared sciatic Nerve Injury (SNI) in control mice (Hif1α-flfl) and in mice with a specific deletion of hypoxia inducible factor 1 alpha (SNS-HIF1α−/−) in DRG neurons. The sciatic nerves were immunostained for the respective redoxins and counterstained with hematoxylin. The redoxin immunoreactivity was quantified with ImageJ. For the DRGs and spinal cord the data show the quantitative assessment of the intensity of redoxin immunoreactivity [1] transformed to rainbow pseudocolors (Fig. 2). In addition, some redoxin examples of the ipsi and contralateral dorsal and ventral horns of the lumbar spinal cord (Fig. 7) and some redoxin examples of the thalamus (Fig. 8) are presented. Characteristics of the antibodies are listed in Table 1 along with some features of the redoxins.
Experimental design, materials and methods
Acknowledgments We acknowledge the financial support of the Deutsche Forschungsgemeinschaft (SFB815 A12 to I.T. and CRC1080 A9). We thank Rohini Kuner for SNScre mice.
Specifications Table Value of the data Data Suppressor of Hairless [Su(H)] is the transcription factor that regulates the expression of the target genes of the Notch signalling pathway [1,2]. Su(H) protein may be phosphorylated by MAPK as a result of Epidermal Growth Factor Receptor (EGFR) activation, providing a means of a direct cross-talk between these two pathways [3–5]. The response of several Notch target genes to the modulations of Su(H) by EGFR signalling activity was analysed by the local overexpression of either wild-type Su(H), a phospho-deficient Su(H) and a phospho-mimetic Su(H) variant [3] using the Gal4::UAS system [6], and staining of the tissues with respective antibodies. Moreover, activated components of the EGFR pathway (DER, rl) were overexpressed alone or in combination with individual Su(H) variants. The response of the Notch target gene cut was observed in cell clones of wing imaginal discs, and the resultant phenotypes on thorax, wings and eyes were recorded in adult flies.
Experimental design, materials and methods