E2F-1 up-regulates TSP1 mRNA expression.

<p>Northern blot analysis of TSP1 expression in 293 cells transfected with HA empty vector of HA-E2F-1 expression vector. β-actin was used as internal control.</p

E2F-1 activates TSP1 promoter.

<p>A, schematic depiction of six TSP1 promoter deletion constructs in the region −2033 to +750bp. The transcription initial site is designated as +1. B, quantification of E2F-1 transcriptional activity on six TSP1 promoter constructs. 293 cells were used for assays and 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. C, dose response of E2F1 on TSP1 promoter activity in U2OS cells. D, the expression of different doses of Flag-tagged E2F1 was verified by Western blot using anti-flag monoclonal antibody, the amounts of Flag-E2F-1 for transfection were indicated. E, the promoter luciferase reporters of E2F1 targeted gene <i>ARF</i>, <i>Apaf1</i> and <i>CyclinE</i> were used as controls to evaluate the transcriptional activity of E2F-1 on TSP1 promoter, 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. F, schematic depiction of four TSP1 promoter deletion constructs in the region −413 to 0 bp. G, quantification of E2F-1 transcriptional activity on four TSP1 promoter constructs, 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. H, the expression of HA-tagged E2F-1 was verified by Western blot using anti-HA monoclonal antibody.</p

E2F-1 directly binds to TSP1 promoter.

<p>A, schematic diagram depicts the region of the TSP1 and actin genes that were amplified. The positions of PCR primers used to detect TSP1 and actin promoter fragments are indicated by arrows. B, 293 cells were treated with formaldehyde to create cross-links between E2F-1 and chromatin. The chromatin was isolated, sheared, and immunoprecipitated (IP) using monoclonal antibody against human E2F-1, or mouse IgG as control. The presence of chromatin fragments corresponding to the <i>TSP1</i> gene or to the <i>β-actin</i> gene promoter was assessed by PCR using gene-specific primers. The gel shows the recovery of TSP1 and actin gene fragments from the protein-chromatin input on the lane 1 (from left to right) as well as those recovered after immunoprecipitation with the anti-E2F-1 antibody (lane 2, up), and with mouse IgG (land 3).</p

E2F-1 binding site in TSP1 promoter is required for E2F-1 up-regulation.

<p>A, the partial sequence of TSP1 promoter (−393 to +27bp). Three potential E2F-1 binding sites were marked by boxes and substitution mutations in those sites were indicated underline. B, schematic of eight TSP1 promoter mutation constructs in the region −393 to 0bp. The three potential E2F-1 binding sites were marked by dark circles and mutations were marked by “X”. C, quantification of E2F-1 transcriptional activity on eight TSP1 promoter mutation constructs, 0.2 µg Flag tagged E2F-1/per well was used for transfection in 24-well plates. D, schematic depiction of E2F-1 DNA binding region (amino acid 110–194) cloned into pVP16 vector. E, quantification of E2F-1 transcriptional activity on three TSP1 promoter mutants, 0.2 µg VP-16E2F-1(110–194) or VP-16/per well was used for transfection in 24-well plates. F, quantification of wild-type E2F-1 and E2F-1(E132) transcriptional activity on TSP1 promoter (−2033–0) luciferase reporter, 0.2 µg vector/per well was used for transfection. G, the expression of HA-tagged wild-type E2F-1 and E2F-1 DNA binding site mutant (E132) was verified by Western blot using anti-Flag monoclonal antibody. H, quantification of pRB influence on E2F-1 transcriptional activity on TSP1 promoter (−2033–0) luciferase reporter construct, the expression of HA tagged human pRB was verified by Western blot and the amounts of HA-pRB for transfection were indicated. I, quantification of E2F-1 deletion mutants on TSP1 promoter (−2033–0) luciferase reporter construct. J, the expression of flag-tagged E2F-1 deletion constructs was verified by Western blot using anti-Flag monoclonal antibody, 0.2 µg different construct/per well was used for transfection. K, quantification of E2F-1 siRNA on TSP1 promoter (−2033–0) luciferase reporter activity. L, the knockdown of E2F1 by siRNA in U2OS cells was verified by Western blot using anti-E2F1 monoclonal antibody, a siRNA targeting GFP was used as control, 0.6 µg siRNA expression vector/per well targeting E2F-1 or GFP was used for transfection in 6-well plates.</p

Installing Logic Gates to Multiresponsive Supramolecular Hydrogel Co-assembled from Phenylalanine Amphiphile and Bis(pyridinyl) Derivative

Recently, logic gates based on multiresponsive hydrogel systems are attractive because of their potential biological applications. A quite simple supramolecular hydrogel co-assembled from phenylalanine-based amphiphile (LPF2) and bis­(pyridinyl) derivative (AP) is constructed. The co-assembled hydrogel exhibited a macroscopic gel–sol transition in response to four distinct input stimuli: temperature, acid, base, and light. A set of techniques including microscopic, spectroscopic, and rheological measurements demonstrate this performance and confirm that the hydrogel is formed through intermolecular hydrogen bonds between amide/pyridine moieties and carbonyl groups. On the basis of its mutiple-stimulus responsiveness, installing gel-based supramolecular logic gates (OR and XOR) is achieved. It may promote the possibility to develop smart soft materials, such as gels, that can be used as tools releasing a drug quantitatively by rational design and fine control of the external stimuli

Influence of C–H···O Hydrogen Bonds on Macroscopic Properties of Supramolecular Assembly

For CH···O hydrogen bonds in assembled structures and the applications, one of the critical issues is how molecular spatial structures affect their interaction modes as well as how to translate the different modes into the macroscopic properties of materials. Herein, coumarin-derived isomeric hydrogelators with different spatial structures are synthesized, where only nitrogen atoms locate at the <i>ortho</i>, <i>meso</i>, or <i>para</i> position in the pyridine ring. The gelators can self-assemble into single crystals and nanofibrous networks through CH···O interactions, which are greatly influenced by nitrogen spatial positions in the pyridine ring, leading to the different self-assembly mechanisms, packing modes, and properties of the nanofibrous networks. Typically, different cell proliferation rates are obtained on the different CH···O bonds driving nanofibrous structures, implying that tiny variation of the stereo-position of nitrogen atoms can be sensitively detected by cells. The study paves a novel way to investigate the influence of isomeric molecular assembly on macroscopic properties and functions of materials

Average Corr and CSI of 3 events for 0–60 min predictions vs. different spatial resolutions of radar using PPLK.

<p>Average Corr and CSI of 3 events for 0–60 min predictions vs. different spatial resolutions of radar using PPLK.</p

Average Corr and CSI of all 8 events for 0–120 min predictions vs. different spatial resolutions of FY-2F using PPLK.

<p>Average Corr and CSI of all 8 events for 0–120 min predictions vs. different spatial resolutions of FY-2F using PPLK.</p

Coefficient of correlation (Corr) and critical success index (CSI) of QPN using Pyramid Lucas-Kanade Optical Flow method (PPLK) vs. 30 min, 60 min, 90 min and 120 min lead time.

<p>for 2338 images of 8 periods using Pixel-based.</p

Generation of Pronounced Resonance Profile of Charge-Transfer Contributions to Surface-Enhanced Raman Scattering

A chemically enhanced mechanism of surface-enhanced Raman scattering (SERS) was investigated using a series of metal-charge-transfer (CT) complex systems fabricated by a self-assembly method. The developed Ag/4-mercaptophenols (MPH)/<i>n</i>-TiO₂ system presented layer number-dependent SERS spectra. By using the electron density values of the Ag₁₃/MPH and Ag₁₃/MPH/TiO₂ system calculated using the density functional theory (DFT) and by using these values in combination with the results of our previous investigations on the mechanism of the Ag/MPH/TiO₂ system, the absorption threshold of the CT complexes was clearly defined. The degree of CT was selected to study the layer number-dependent SERS spectra. Based on the layer number-dependent SERS data, it has been inferred that the degree of CT represents a resonance phenomenon. In addition, the CT resonance occurs at higher energy in the Ag/MPH/<i>n</i>-TiO₂ system than in the monolayer TiO₂ system owing to the blue-shift of CT states with the continuous introduction of TiO₂. Thus, we provide a good example of the use of a CT complex system to investigate the chemical mechanism of SERS