br Experimental design materials and methods br

Experimental design, materials and methods
Acknowledgements The authors are grateful for a DFG Grant (German Research Council, graduate school GRK1505/2 \"WELISA\"), which covered the expenses for consumables as well as the Ph.D. position of A. Galow. We wish to thank R. Modrozynski for his technical assistance. R. Sleigh is acknowledged for his help with the language.
Experimental design, materials and methods
Data This data describes the proteome of subcutaneous adipose tissues obtained from late pregnant dairy cows during summer heat stress or during the winter season. Adipose tissue has a central role in the regulation of metabolism in dairy cows, and many proteins expressed in this tissue are involved in metabolic responses to stress (Peinado et al., 2012) [1]. Environmental heat stress is one of the main stressors limiting production in dairy cattle (Fuquay, 1981; West, 2003) [2,3], and there is a complex interaction between heat stress and the transition period from late pregnancy to onset of lactation, which is manifested in heat-stressed late-gestation cows (Tao and Dahl, 2013) [4].Table 1 contains the list of 1495 identified and quantified proteins obtained by proteomic analysis.
Acknowledgments The authors wish to thank the Volcani dairy farm staff for their assistance with animal care. This research was supported by a Grant from the GIF, the German-Israeli Foundation for Scientific Research and Development, Young Scientist Grant [Grant number I-2403-204.12/2015]. G.F. is the Incumbent of the David and Stacey Cynamon Research Fellow Chair in Genetics and Personalized Medicine.
Data The extracts of human liver or HepG2 vesicular monoamine transporter were treated with trypsin using the FASP protocol. The peptides produced were analyzed by ESI LC-MS/MS. The lists of proteins detected are presented in Supplementary Table 1. The extracts of human liver tissue (300µg of protein) were also run by 2-DE (Fig. 1). The gel produced was stained with Coomassie Brilliant Blue R350. Image analysis was performed by ImageMaster 2D Platinum software (GE Healthcare, Pittsburgh, PA, USA). Next, 96 sections were selected, given pI/Mw coordinates, and cut for subsequent ESI LC-MS/MS analysis (Fig. 1). A list of all proteins detected by Mascot (only without hemoglobin) in the human liver extracts is presented in Supplementary Table 2. Hemoglobin was removed as a major contaminant of blood plasma proteins. If the same protein was identified in different sections, it was considered to exist as different proteoforms. According to this rule, a total of 14667 proteoforms were identified.
Experimental design, materials and methods
Acknowledgements The study was supported by Russian Scientific Foundation, grant # 15–15-30041. We acknowledge the IBMC \"Human Proteome\" Core Facility for assistance with the generation of mass-spectrometry data. Carita Lanner is acknowledged for the editing assistance. All the authors declare that they have no conflict of interest.
Data The dataset of this article provides information on GALR3 mutants that stabilize the receptor in either an agonist-bound or antagonist-bound form. Table 1 shows how 23 of the 210 point mutants expressed on VLPs increase [125I]-galanin binding and thermal stability compared to WT. In addition, a direct correlation of the Bmax values with recombinant expression yields of the mutants in Sf9 cultures was shown (Table 1 and Fig. 1). Combinations of these mutants were made to find the best thermostabilizing GALR3 agonist-bound (Table 2) and GALR3 antagonist-bound (Table 3) variants.
Experimental design, materials and methods
Acknowledgements
Data A new probe to detect sulfatase activity was developed. When the probe was treated with sulfatase, it generated fluorescent N-methylisoindole which was influenced by reducing agents and pH conditions [1]. The fluorescence intensity, kinetic parameters and inhibitory potency under Tris buffer and reducing agents were described. Fluorescence intensities of the probe with sulfatase at pH 5 and 7 were compared.