br Acknowledgments We would like to

Acknowledgments We would like to thank Dr. Daniel Conrad and Julie Farnsworth at the Flow Cytometry Research Core at the Massey Cancer Center, VCU. This project was funded by the National Institute of Justice Award number 2013-DN-BX-K033.
Data
Experimental design, materials and methods
Acknowledgments The work received support from NHLBI for 1RO1 HL6976-01 to Dr. Mary Taub.
Data We provide additional data about the stability and functionality of nAChR solubilized from Torpedo californica with the lipid analog detergent, 1-hexadecanoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (LFG-16). The stability of the affinity purified nAChR-LFG-16 detergent complex was determined after it incorporation into lipid cubic phase (LCP) of 1-(cis-9-Octadecenoyl)-rac-glycero for a period of 30 days using Fluorescence Recovery after Photobleaching (FRAP)(Fig. 1). The functionality of the purified nAChR-LFG-16 detergent complex was studied after reconstitution into Xenopus what is doxycycline hyclate by mean of two electrode voltage clamp (Fig. 2).
Experimental design, materials and methods
Acknowledgments This research was supported by the National Institutes of Health NIGMS Grants 1R01GM098343, the RISE Program (2R25GM061151), MARC Program (5T34GM007821) and Neuroimaging and Electrophysiology Facility Grant NIH P20 GM 103642. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Data BAFF and APRIL belong to a family of closely related TNF family ligands [1,2]. Although crystal structures of BAFF or APRIL homotrimers are known since several years, we only recently reported the crystal structure of BAFF and APRIL heterotrimers [3]. In order to generate a homogeneous protein material for structural studies, we joined one APRIL and two BAFF subunits into a single chain protein, by introducing two glycine-serine linkers (GGGGS) in between ligand subunits. The expressed protein APRIL–BAFF–BAFF was crystalized and its X-ray diffraction structure was solved and deposited into Protein Data Bank with accession code 4ZCH [3].
Experimental design, materials and methods
Crystallization The purified protein was crystallized by trying various pH conditions and other crystallization factors. The hit conditions were optimized to obtain crystals suitable for X-ray diffraction measurements. Crystals were obtained by the hanging drop vapor diffusion method incubated at 20°C. Protein solution at 6mg/ml in 20mM Hepes/NaOH pH 7.5, 150mM NaCl (0.5µl) was mixed with 0.5μl of a reservoir solution of 0.1M Tris/HCl, pH 8.75, 14% PEG6000 (w/v), 1M LiCl in a 1-to-1 ratio. Once obtained, crystals were mixed with reservoir solution supplemented with 10% (v/v) 2,3-butanediol prior to flash freezing in liquid nitrogen.
Data collection and processing The diffraction data were collected at 100°K at X-ray wavelength of 0.99998 Å at beamline X06SA/Swiss Light Source (SLS) using a Pilatus 6M detector, and integrated using the software XDS and XSCALE [4]. Table 1 shows the parameters used in the data collection. The crystal system was determined to be orthorhombic with space group C2221. Its unit cell dimensions were of 57.04Å, 117.86Å and 295.52Å. Data were processed to 2.43Å resolution. A total of 134,837 reflections were measured, referring to 36,901 unique reflections, representing a completeness of 96.7% and a redundancy of 3.7. The average signal to noise ratio was 13.46 for the whole data set and 3.06 for the highest resolution shell (2.68–2.43Å). The data set quality is further assessed by two quantities, Rsym and Rmerge, in order to measure internal agreement (residual factors) of symmetry-related reflections and redundant data. The Rsym and Rmerge were 7.9% and 9.2%, respectively. Table 2 shows the correlation between observed and expected profiles, and Tables 3 and 4 show the R-factors and Wilson statistics of the data set, respectively.