Coefficient matrix among the samples in RNA-Seq from bovine mammary epithelial cells stimulated by ten different essential amino acids

Coefficient matrix among the samples in RNA-Seq from bovine mammary epithelial cells stimulated by ten different essential amino acids</p

Non-targeted metabolomics data from bovine mammary epithelial cells stimulated by ten different essential amino acids

Non-targeted metabolomics data from bovine mammary epithelial cells stimulated by ten different essential amino acids.</p

Sequences of the truncated NS1-27 polypepide.

<p>Sequences of the truncated NS1-27 polypepide.</p

Primary screening of epitope with mAb 3G2.

<p>One 16-AA polypeptide of TMUV NS1 protein (NS1-27) was screened with mAb 3G2 by indirect ELISA. Mouse serum against TMUV NS1 protein and normal mouse serum were used as positive and negative controls, respectively. Each sample was detected in triplicate. Error bars were expressed as standard deviation of the means (n = 3). The mean value was statistically significant, calculated by the two-tailed Student’s unpaired t-test (*P < 0.05).</p

IFA and western-blot identification of mAb 3G2.

<p>A: Western-blot identification Lane 1, Control, GST-tag didn’t react with mAb 3G2; Lane 2, The band of NS1-GST fusion protein was reacted with mAb 3G2; Lane M, Blue plusIIprotein Marker (14-120kda, Transgen Biotech). B: IFA identification Monoclonal antibody against TMUV NS1 protein was used to perform IFA on TMUV-infected BHK-21 cells. BHK-21 cells infected with TMUV yielded significant fluorescence with six MAbs in the cytoplasm; Control BHK-21 cells didn’t yield any fluorescence.</p

Western-blot identification of epitope.

<p>The 16-AA polypeptide of NS1-27 reacted with mAb 3G2 in Western-blot assay. Lane M, PageRuler Prestained Protein Ladder (Fermentas, Canada); Lane 1, GST-tag didn’t react with mAb 3G2; Lane 2, The band of NS1-27-GST fusion protein was visualized with mAb 3G2.</p

Sequences of the overlapping polypeptides from TMUV NS1 (SDSG strain, Accession number: KJ740747.1).

<p>Sequences of the overlapping polypeptides from TMUV NS1 (SDSG strain, Accession number: KJ740747.1).</p

The accurate mapping of one B cell epitope with mAb.

<p>NS1-27 polypeptide was truncated from the carboxy and amino terminals. After the truncated peptides were expressed as a GST fusion protein, they were probed with mAb 3G2 by indirect ELISA respectively. The minimal unit of the peptide was the sequence of 8 AA and 3 AA truncated from the carboxy and amino terminals of NS1-27.</p

Templated Biomineralization on Self-Assembled Protein Nanofibers Buried in Calcium Oxalate Raphides of <i>Musa</i> spp.

Biological organisms possess an unparalleled ability to control crystallization of biominerals with convoluted internal structures. For example, an occluded organic matrix can interact with the mineral during its formation to control its morphology and structure. Although related matrix proteins that preferentially nucleate minerals have been identified, the mechanisms elucidating the structural and chemical complexity of calcium oxalate biominerals in plants remain unclear. Here, we show that a protein nanofiber (14 kDa) is embedded inside raphide (needle-shaped calcium oxalate) crystals of banana (<i>Musa</i> spp.), and that nanometer-scaled calcium oxalate spheres are arranged along the long axes of this central proteinaceous filament to form laminated structures through an aggregation-based growth mechanism, resulting in the final product of elongated and tapered hexagonal crystals. We further demonstrate that 11 amino acid peptide segments, with hydrophilic and hydrophobic residues rich in proline derived from the C-terminus of this full protein sequence, in vitro self-assemble into fibers and accelerate calcium oxalate nucleation kinetics. Remarkably, elongated and organized microstructures which are similar in appearance to natural raphide crystals are formed, emphasizing interactions between the mineral and self-assembled protein fibers. We anticipate that the present investigation of the structural and morphological complexity of plant calcium oxalate crystals and the underlying mechanisms of their formation will contribute to our understanding not only how plants evolved these sophisticated structures and morphologies for survival and adaptation, but also ultimately provide useful clues about how to maximally sequester calcium ions and/or oxalate in a confined compartment

Tuning Biocompatible Block Copolymer Micelles by Varying Solvent Composition: Dynamics and Populations of Micelles and Unimers

Optimization of micellar molecular encapsulation systems, such as drug delivery vehicles, can be achieved through fundamental understanding of block copolymer micelle structure and dynamics. Herein, we present a study of PEO–PCL block copolymer spherical micelles that self-assemble at 1% wt/vol in D₂O–THF-<i>d</i>₈ mixtures. Varying solvent composition as a function of cosolvent THF-<i>d</i>₈ at constant polymer concentration (1% wt/vol) allows sensitive study of how small molecule additives influence micelle structure and dynamics. We conduct nuclear magnetic resonance spectroscopy and diffusometry on two block copolymer (2k series: PEO_2k–PCL_3k; 5k series: PEO_5k–PCL_8k) spherical micelles that show drastically different behaviors. Unimers and micelles coexist in solution from 10–60 vol % THF-<i>d</i>₈ for the 2k series but only coexist at 60 vol % THF-<i>d</i>₈ for the 5k series. At ≥ 60 vol % THF-<i>d</i>₈ micelles disassemble into free unimers for both series. We observe relatively flat micelle diffusion coefficients (∼1 × 10^–10 m²/s) with increasing THF-<i>d</i>₈ below 60 vol % for both 2k and 5k series, with only small changes in micelle hydrodynamic radius (≈14 nm) over this range. We compare these results to a detailed SANS and microscopy study described in a companion paper. These fundamental molecular dynamics, unimer population, and diffusion results, as a function of polymer composition and solution environment, provide critical fodder for controlled design of block copolymer self-assembly