MOESM1 of Construction of a shuttle expression vector for lactic acid bacteria

Additional file 1: Figure S1. Agarose gel electrophoresis of various plasmids used in the study, (a) Lane 1: NEB Supercoiled DNA ladder; Lane 2: Shuttle vector pPBT-GFP; Lane 3: Plasmid pLES003; Lane 4: Plasmid pCP289; Lane 5: Vector pMK-RQ. Multiple bands are visible due to supercoiled, partially nicked and linear forms of plasmids. Figure S2. Agarose gel electrophoresis of DNA fragments used for vector development, Lane 1: Novagen Perfect DNA ladder; Lane 2: Pediocin operon; Lane 3: oriLB; Lane 4: oriPA; Lane 5: bsh; Lane 6: gfp. Sequence of Shuttle Vector pPBT-GFP, 9.6 kb

Number of Transcription Factor binding sites on different A-P patterning genes in <i>Drosophila melanogaster</i> and <i>Tribolium castaneum</i> as predicted by the MCAST software.

Here, Drosophila melanogaster and Tribolium castaneum are abbreviated as Dm and Tc respectively.</p

Exploiting the Biophysical Cues toward Dual Differentiation of hMSC’s within Geometrical Patterns

A wide variety of cues from the extracellular matrix (ECM) have been known to affect the differentiation of stem cells in vivo. In particular, the biophysical cues and cell shape have been known to affect the stem cell function, yet very little is known about the interplay between how these cues control differentiation. For the first time, by using photolithography to pattern poly(ethylene glycol) (PEG), patterns of square and triangular geometries were created, and the effect of these structures and the biophysical cues arising were utilized to differentiate cells into multiple lineages inside a same pattern without the use of any adhered protein or growth factors. The data from these studies showed that the cells present at the edges were well elongated, exhibit high aspect ratios, and differentiated into osteogenic lineage, whereas the cells present at the center exhibit lower aspect ratio and were primarily adipogenic lineage regardless of the geometry. This was correlated to the higher expression of focal adhesion proteins at the edges, the expression of which have been known to affect the osteogenic differentiation. By showing MSC lineage commitment relationships due to physical signals, this study highlights the importance of these cues and cell shape in further understanding stem cell behavior for tissue engineering applications

Multiporphyrin Arrays on Cyclotriphosphazene Scaffolds

We report the synthesis of first examples of hexaporphyrin and dodecaporphyrin assemblies on cyclotriphosphazene scaffold by adopting two different approaches based on Ru-pyridyl “N” coordination in decent yields. The multiporphyrin assemblies were confirmed by ³¹P, ¹³C, ¹H, ¹H–¹H COSY, and NOESY NMR spectroscopic studies. The absorption studies showed 2-fold intensity enhancement with negligible changes in peak maxima compared to porphyrin monomers. The redox potentials of multiporphyrin assemblies showed the redox features of the constituted porphyrin monomers and supported weak interactions among the porphyrin units in noncovalent hexaporphyrin and dodecaporphyrin arrays

List of genes used for enhancer localization along with their chromosomal locations, accession numbers, gene locations, and gene sequence location with the flanking regions in <i>Drosophila melanogaster</i>.

List of genes used for enhancer localization along with their chromosomal locations, accession numbers, gene locations, and gene sequence location with the flanking regions in Drosophila melanogaster.</p

Schematic representation of the regulatory relationship between the anterior-posterior patterning gene cascade in <i>Drosophila melanogaster</i>.

Schematic representation of the regulatory relationship between the anterior-posterior patterning gene cascade in Drosophila melanogaster.</p

Location of the transcription start sites and putative cis-regulatory modules and predicted CRM’s size as predicted by the MCAST software on the A-P patterning genes of <i>Tribolium castaneum</i>.

Location of the transcription start sites and putative cis-regulatory modules and predicted CRM’s size as predicted by the MCAST software on the A-P patterning genes of Tribolium castaneum.</p

Location of the transcription start sites and putative cis-regulatory modules and CRM’s size as predicted by the MCAST software on the A-P patterning genes of <i>Drosophila melanogaster</i>.

Here–and + represents the upstream and downstream location of predicted CRM respectively.</p

3D Bioprinted Alginate-Silk-Based Smart Cell-Instructive Scaffolds for Dual Differentiation of Human Mesenchymal Stem Cells

Designing smart bioinks, which can provide multifunctionality and instructive cues to cells, is a current need of the tissue engineering field. Addressing these parameters, this work aims at developing a smart dual 3D bioprinted scaffold that is capable of differentiating human mesenchymal stem cells into two different lineages within the same construct without providing any exogenous cues. Here, biocompatible alginate- and silk-based bioinks were developed to print self-standing structures with the ability of spatially controlled differentiation of the encapsulated hMSCs. We present this proof of concept and have demonstrated a smart design where the incorporation of phosphate groups enhanced the osteogenic differentiation, whereas the addition of silk promoted the chondrogenic differentiation. Altogether, the present work suggests the potential of the developed bioinks for use in creating clinically viable osteochondral grafts

List of genes used for enhancer localization along with their chromosomal locations, accession numbers, gene locations, and gene sequence location with the flanking regions in <i>Tribolium castaneum</i>.

List of genes used for enhancer localization along with their chromosomal locations, accession numbers, gene locations, and gene sequence location with the flanking regions in Tribolium castaneum.</p