Knockdown of IGF-1R/INSR.

<p>Control or INSR siRNA were transfected into hTCEpi cells. Two to three days after transfection, WCL, cytosolic (Cyto) and nuclear fractions (Nuclear) were collected. (A) Immunoblots of WCL with antibodies against INSR (C-19), IGF-1R (CST#3027), or β-actin (loading control). (B) WCL of hTCEpi cells were immunoprecipitated with antibodies αIR3, C-19, or MA-20 (against INSRα and only reacts with INSR/INSR). Immunoprecipitates were then immunoblotted with anti-IGF-1Rβ (CST#3027) or anti-INSRβ (C-19). (C) Cytosolic and nuclear fractions of hTCEpi cells were subjected to immunoprecipitation with antibodies αIR3 or C-19. Immunoprecipitates were then immunoblotted with anti-IGF-1Rβ (CST#3027). GAPDH is the marker for cytosolic extract; and SP1 is the marker for nuclear extract.</p

Presence of IGF-1 receptor and insulin receptor (IGF-1R/INSR) hybrid in corneal epithelium.

<p>(A) Reciprocal immunoprecipitation (IP) of IGF-1 receptor and insulin receptor in the hTCEpi cell line. Whole cell lysates were subjected to IP with antibodies against either the IGF-1 receptor (IGF-1R) β-subunit (CST#3027) or the insulin receptor (INSR) β-subunit (C-19). Immunoprecipitated products were then immunoblotted with the same antibodies. Input represents the whole cell lysates before IP. (B) Co-precipitation of IGF-1R/INSR hybrid in primary corneal epithelial cell cultures. Protein lysates from passage 3 of human corneal epithelial cells were equally immunoprecipitated with anti-IGF-1Rβ (CST#3027) or anti-INSRβ (C-19), and detection of IGF-1R/IGF-1R or IGF-1R/INSR was performed with immunoblots with the same antibody against IGF-1Rβ.</p

Electrospun Nanofibrous Composite Membranes for Separations

ConspectusElectrospinning is regarded as an efficient method for directly and continuously fabricating nanofibers. The electrospinning process is relatively simple and convenient to operate and can be used to prepare polymer nanofibers for almost all polymer solutions, melts, emulsions, and suspensions with sufficient viscosity. In addition, inorganic nanofibers can also be prepared via electrospinning by adding small amounts of polymers into the inorganic precursors, which are generally regarded as nonspinnable. The diameter of the electrospun nanofibers can be tuned from tens of nanometers to submicrons by changing the spinning parameters. The nonwoven fabric stacked with electrospun fibers is a porous material with interconnected submicron pores, providing a porosity above 80%. However, limited by the unstable rheological properties of the electrospinning fluid, it is difficult to obtain nanofibers stably and continuously with an average diameter of <100 nm, which narrows the separation applications of the electrospun nanofibrous membranes to only microfiltration, air filtration, or use as membrane substrates. Therefore, to fully take advantage of electrospun nanofibrous membranes in other separation applications, electrospun nanofibrous composite (ENC) membranes were developed to improve and optimize their selectivity, permeability, and other separation performances. The composite membranes not only have all the advantages of single-layered or single-component membranes, but also have more flexibility in the choice of functional components.In this account, we summarize the two combination strategies to design and fabricate ENC membranes. One is based on the component combination, in which functional components are homogeneously or heterogeneously mixed in the fiber matrix or modified on the nanofiber surface. The other one is termed as the interfacial combination, in which functional skin layers are fabricated on the top of the electrospun membranes via interfacial deposition or interfacial polymerization, to construct selective barriers. The specific preparation approaches in the two combination strategies are discussed systematically. Additionally, the structural characteristics and separation performances of ENC membranes fabricated via these approaches are also compared and analyzed to clarify their advantages and range of utilization. Subsequently, the six applications of ENC membranes we focus on are demonstrated, including adsorption, membrane distillation, oil/water emulsion separation, nanofiltration, hemodialysis, and pervaporation. To meet their different requirements for separations, our consideration about the choice of combination strategies, related preparation methods, and functional components are discussed based on typical research cases. In the end, we conclude this account with an overview of the challenges in industrial manufacturing, mechanical strength, and interfacial attachment of ENC membranes and prospect their future developments

Gene ontology clustering of gene annotations.

<p>The enriched terms associated with the smallest p-value and with p<0.1 were shown.</p>a<p>original database where the terms orient.</p>b<p>genes involved in the term.</p>c<p>percentage  =  involved genes/total genes.</p>d<p>modified Fisher Exact p-value is adopted to measure the gene-enrichment in annotation terms.</p

GeneSolution^a siRNA target sequences.

a<p>GeneSolution contains equal mixture of four siRNA duplexes.</p

Subcellular localization of IGF-1R/IGF-1R and IGF-1R/INSR hybrid.

<p>(A) Presence of IGF-1R/INSR hybrid in the nucleus. Cytosolic (Cyto) and nuclear (Nuclear) fractions of hTCEpi cells were immunoprecipitated with anti-IGF-1Rβ (CST#3027) or anti-INSRβ (C-19) and then immunoblotted with the same antibodies. Bottom left: Immunoblot of IGF-1R immunoprecipitates with anti-INSR indicated the presence of hybrid in the nuclear fraction. (B) Exclusive presence of hybrid in the nucleus. Whole cell lysates (WCL) and nuclear fractions (Nuclear) of hTCEpi cells were subjected to immunoprecipitation with antibodies against IGF-1Rα (αIR3, which reacts with IGF-1R/IGF-1R) or INSRβ (C-19, which reacts with IGF-1R/INSR and INSR/INSR). Immunoprecipitates were then immunoblotted with anti-IGF-1Rβ. Hybrid-R was shown in the WCL and nuclear fractions but IGF-1R/IGF-1R was only detected in the WCL. Controls for cross-contamination between each compartment were confirmed by immunoblotting with antibodies against GAPDH (cytosolic extract) and SP1 (nuclear extract).</p

Functional clustering of gene annotations using the DAVID resource.

<p>Functional clustering of gene annotations using the DAVID resource.</p

Characteristic Swelling–Deswelling of Polymer/Clay Nanocomposite Gels

Swelling behavior of nanocomposite hydrogels (NC gels) having organic polymer/inorganic clay network structures were systematically investigated, focusing on the role of exfoliated clay platelets with sodium counterions in the network and effects of various swelling conditions. NC gels in water exhibited characteristic swelling–deswelling behavior, i.e., initial large swelling, maximum swelling, and subsequent deswelling toward an equilibrium state, under conditions where the water was changed frequently. Effects of swelling conditions, such as the frequency of changing the water, amount of water per unit gram of gel, salt concentration, and pH of the swelling solvent, gel composition (e.g., kind of polymer, clay concentration (<i>C</i>_clay), and polymer concentration (<i>C</i>_p)), swelling temperature, and gel size, were clarified. NC gels with different polymers were all found to exhibit swelling–deswelling behavior except at very low <i>C</i>_clay and high <i>C</i>_p. Spontaneous deswelling of the gels was attributed to the combined effects of high swelling capability of the NC gel as a polyelectrolyte gel and continuous release of sodium ions from the network during swelling. Furthermore, the swelling–deswelling behavior could be reversed by reintroducing sodium ions into the network. These characteristic swelling behaviors of NC gels with polymer/clay networks are completely different from those of hydrogels with chemically cross-linked polymer network structures

IGF-1 activates the IGF-1R/INSR hybrid.

<p>(A) Existence of IGF-1R and INSR αβ-dimers after reducing immunoprecipitation. hTCEpi lysates were reduced with DTT to break the receptors into αβ-dimers. Reduced lysates were then immunoprecipitated with rabbit polyclonal anti-IGF-1Rβ (CST#3027) or anti-INSRβ (C-19). <i>Top panel</i>: Immunoblot of INSRβ immunoprecipitates from reduced lysates with anti-IGF-1Rβ showed no or faint bands corresponding to the IGF-1Rβ. <i>Bottom panel</i>: Immunoblot of IGF-1Rβ immunoprecipitates from reduced lysates with anti-INSRβ showed no or faint bands corresponding to the INSRβ. (B) Activation of IGF-1R and INSR in hTCEpi cells by IGF-1 stimulation. Protein lysates of hTCEpi cells after stimulation by IGF-1 (100 ng/ml) or insulin (100 ng/ml) for 15 min were collected and reduced by DTT and then subjected to reducing IP. IGF-1, but not insulin, induced phosphorylation of IGF-1 receptor (<i>Top panel</i>) and insulin receptor (<i>Bottom panel</i>) detected on immunoblots with anti-PY20.</p

Poly(amidoamine) Dendrimer-Enabled Simultaneous Stabilization and Functionalization of Electrospun Poly(γ-glutamic acid) Nanofibers

We report a facile and general approach to using generation 2 (G2) poly­(amidoamine) (PAMAM) dendrimers for simultaneous stabilization and functionalization of electrospun poly­(γ-glutamic acid) nanofibers (γ-PGA NFs). In this study, uniform γ-PGA NFs with a smooth morphology were generated using electrospinning technology. In order to endow the NFs with good water stability, amine-terminated G2.NH2 PAMAM dendrimers were utilized to crosslink the γ-PGA NFs via 1-ethyl-3-(3-dimethylami-nopropyl) carbodiimide coupling chemistry. Under the optimized crosslinking conditions, G2.NH2 dendrimers partially modified with fluorescein isothiocyanate (FI) or folic acid (FA) were used to crosslink γ-PGA NFs. Our results reveal that G2.NH2–FI is able to simultaneously render the NFs with good water stability and fluorescence property, while G2.NH2–FA is able to simultaneously endow the NFs with water stability and the ability to capture FA receptor-overexpressing cancer cells in vitro via ligand–receptor interaction. With the tunable dendrimer surface chemistry, multifunctional water-stable γ-PGA-based NFs may be generated via a dendrimer crosslinking approach, thereby providing diverse applications in the areas of biosensing, tissue engineering, drug delivery, and environmental sciences