br Acknowledgments The SAXS data collection was

Acknowledgments The SAXS data collection was done at the DND-CAT at the Advanced Photon Source Science User Facility operated for the U.S. Department of Energy (DOE), supported by the U.S. DOE under Contract no. DE-AC02-06CH11357. This project has been funded with Federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under Contract nos. HHSN272200700058C and HHSN272201200026C (WFA), and the National Science Foundation Grant MCB 1024945 (MAB).
Data
The recombinant human mAb (IgG1) and IgG1-fusion protein were manufactured using standard cell culture, purification and formulation processes [2]. The IgG1 has two identical light chains and two identical heavy chains with molecular weight of 150kDa and N-glycosylation site at N297. The IgG fusion protein, approximately 90kDa, was comprised of two chains each with a human protein domain fused onto the N-terminus of a human IgG1 constant domain (Fc). The protein has three N-glycosylation sites at N76, N108 and N207.
Analysis of the IgG protein The IgG protein was analyzed 1) in reduced form by MALDI-ISD in a 12T Fourier Transform Ion Cyclotron Resonance (FT-ICR XR) mass spectrometer (Bruker, Bremen, Germany), and 2) after IdeS digestion and dithiothreitol (DTT) reduction to generate three smaller fragments; light chain, Fab, and Fc/2 after which analyzed by LC–MS/MS on an Orbitrap Elite (Thermo Scientific, Saint Jose, CA, USA). Intact protein mass measurement for detection of protein species of the IgG was performed using ESI FT-ICR MS as demonstrated in the related study [1].
Analysis of the IgG fusion protein
Acknowledgments The data collection was supported in part by collaborative grant from MedImmune, LLC (2014-2312 RSA), and University of Maryland, Baltimore, School of Pharmacy Mass Spectrometry Center (SOP1841-IQB2014).
Value of the data
Data
Experimental design, materials and methods
Data Mean values for number and area of the focal adhesion points of MC3T3-E1 Nanaomycin A on the PDMS substrates, amount of vinculin based on analysis of images from immunofluorescent labeling for each PDMS substrate of varying stiffness and loading (cyclic or static) condition were obtained based on two experimental runs. Substrate stiffness, expressed as the modulus of elasticity, was determined for the PDMS substrates used in the culture chambers through standard mechanical testing. The values presented are mean values obtained from five different measurements using linear regression on their respective engineering stress-strain curves. Mean and standard errors, for each substrate׳s modulus of elasticity, “E”, are as follows: 2.04±0.06MPa for PDMS 5, 1.70±0.05MPa for PDMS 10 and 1.22±0.05MPa for PDMS 15.
Conflicts of interest
Acknowledgments The authors acknowledge the support provided through NIH Grant no. R25GM088023 and HHMI Grant no. 52007566. Jose Almodovar from the University of Puerto Rico- Mayagüez Campus assisted in the acquisition of the immunofluorescence images.
Data To identify the differentially expressed genes in the fission yeast under nitrosative stress, cells were treated with two different doses of pure NO donor i.e. 0.1mM and 1mM for 15min. It was previously reported that fission yeast Schizosaccharomyces pombe cells are much more sensitive to a concentration of 3mM of detaNONOate than to 1mM [1,2] in terms of cell growth, lowering the mitotic index, while cell viability is maintained at 95%. The differentially expressed genes are listed in Table 1. Treatment of the wild type S. pombe cells with NO donor compound detaNONOate at a concentration of 1mM resulted in 9 genes to be repressed while 30 genes were identified as induced (Table 2). The differential expressions of 8 genes were confirmed by Real-Time (RT) PCR analysis. The gene expression profiles (up regulation or down regulation) obtained by differential display analysis and by RT-PCR are similar as listed in Table 3. Gene Ontology (GO) terms that were enriched in the differentially expressed gene lists were searched using the online tool DAVID. In order to identify the different pathways affected under nitrosative stress, the information provided in KEGG was referred. Genes were classified as belonging to the different pathways that were affected upon treatment with pure NO donor compound. Fig. 1 shows the 18 pathways that were significantly affected (p<0.01) when the wild type S. pombe cells were treated with 0.1mM concentration of pure NO donor detaNONOate for 15min at 30°C. Fig. 2 shows the 24 pathways that were significantly affected (p<0.01) when the cells were treated with 1mM concentration of pure NO donor under similar conditions.