Reactive Dendronized Copolymer of Styryl Dendron and Maleic Anhydride: A Single Molecular Scaffold

Novel dendronized copolymers bearing reactive anhydride groups along the backbones are reported in this paper. They were synthesized through copolymerizations of styryl macromonomers bearing Fréchet-type dendrons of the first to the fourth generation and maleic anhydride (MAn) through conventional radical polymerization. The dendritic macromonomers were prepared by an accelerated convergent approach, i.e., a reaction of the dendritic bromide of lower generation with a styrene derivative bearing a 3,5-dihydroxybenzyl group. The dendronized copolymers with rather high molar masses were obtained under mild conditions, even for the fourth-generation dendritic monomer as determined by static light scattering (SLS). For example, the degree of copolymerization for the third-generation monomer reached 487. Owing to the reactivity of the anhydride, functional groups, which were buried by the grafted dendrons along the backbone, could be easily introduced as follows:  (1) by the hydrolysis under acid conditions, the amphiphilic copolymers with a structure of dendron-alt-2COOH were prepared; (2) by reacting with alkyl primary amine, the copolymers with dendron-alt-linear alkyl chain were obtained in a quantitatively yields; (3) when reacted with a primary amino dendron of its generation different from the pendent dendron, a series of new dendronized copolymers with their side dendrons grafted in an alternative generation structure were produced. Therefore, the novel reactive dendronized copolymer presented herein can be a single molecular scaffold to be applied in nanomaterials and nanotechnologies of one dimension

Ectopic expression of mutant p53 R163H cooperates with p53-KD to alter cyst morphology.

<p><b>A</b>, Generation of MCF-10A cell lines in which siRNA-resistant mutant p53-R163H was expressed along with knockdown of endogenous wild-type p53. The levels of wide-type p53 and mutant p53-R163H were determined by Western blotting. <b>B</b>, The level of wild-type p53 transcripts was determined by RT-PCR. <b>C</b>, Representative images of MDCK cells or MDCK cells with p53-KD-R163H in 2-D culture. <b>D</b>, Representative images of MDCK cells or MDCK cells with wild-type p53-KD and overexpression of mutant p53-R163H in 3-D culture for 12 d. Scale bar: 100 µM. <b>E</b>, Top panel: colony formation assay was performed with MDCK cells or MDCK cells with p53-KD and overexpression of R163H. Bottom panel: the number of colonies was counted and presented as Mean ± SD from three separate experiments. <b>F</b>, Wound healing assay was performed with MDCK cells, MDCK cells with p53-KD, or MDCK cells with p53-KD and overexpression of R163H. Top panel: cell migration was determined by visual assessment of cells migrating into the wound for 24 h using a phase-contrast microscopy. Bottom panel: the time required for wound closure was measured and presented as mean ± SD from three separate experiments.</p

EMT markers are regulated upon ectopic expression of mutant p53, some of which are further enhanced by knockdown of endogenous wild-type p53 in MDCK cells.

<p><b>A</b>-<b>C</b>, Western blots were prepared with extracts from parental MDCK cells (lane 1), p53-KD MDCK cells (lane 2), MDCK cells in which a mutant p53 was ectopically expressed (lanes 3, 5) and MDCK cells in which a mutant p53 was ectopically expressed along with knockdown of endogenous wild-type p53 (lanes 4, 6). The blots were probed with antibodies against β-catenin (A), E-cadherin (A), Snail (B), Slug (B), Twist (B), c-Met (C) and actin (A-C). The protein levels of EMT markers were quantified and the ratios were labeled under the corresponding bands. <b>D</b>, Proposed model of mutant p53 in MDCK cell tubulogenesis. </p

Overexpression of mutant p53-R261H disrupted tubular formation in 3-D culture.

<p><b>A</b>, Generation of MDCK cell lines in which siRNA-resistant mutant p53-R261H was stably overexpressed (clones 1 and 2). The protein levels of mutant p53-R261H and actin were measured by Western blotting. <b>B</b>, The level of wild-type p53 transcripts was determined by RT-PCR. <b>C</b>, Representative images of MDCK cells, MDCK cells with p53 knockdown, or MDCK cells with mutant p53-R261H in 2-D culture (200×). <b>D</b>, Representative images of MDCK cells with mutant p53-R261H in 3-D culture. Scale bar: 100 µM. <b>E</b>, Top panel: colony formation assay was performed with MDCK cells or MDCK cells with mutant p53-R261H. Bottom panel: the number of colonies was counted and presented as Mean ± SD from three separate experiments. <b>F</b>, Wound healing assay was performed with MDCK cells, MDCK cells with p53-KD, or MDCK cells with mutant p53-R261H. Top panel: cell migration was determined by visual assessment of cells migrating into the wound for 24 h using a phase-contrast microscopy. Bottom panel: the time required for wound closure was measured and presented as mean ± SD from three separate experiments.</p

Overexpression of mutant p53 R163H disrupted tubular formation in 3-D culture.

<p><b>A</b>, Generation of MDCK cell lines in which siRNA-resistant mutant p53-R163H was stably overexpressed (clones 3 and 5). The level of p53-R163H was determined by Western blotting. <b>B</b>, The level of wild-type p53 transcripts was determined by RT-PCR. <b>C</b>, Representative images of MDCK cells, MDCK cells with p53 knockdown, or MDCK cells with mutant p53 (R163H) in 2-D culture (200×). <b>D</b>, Representative images of MDCK cells, MDCK cells with p53 knockdown, or MDCK cells with mutant p53-R163H in 3-D culture for 6 d or 12 d. Scale bar: 100 µM. <b>E</b>, Top panel: colony formation assay was performed with MDCK cells, MDCK cells with p53 knockdown, or MDCK cells with mutant p53-R163H. Bottom panel: the number of colonies was counted and presented as Mean ± SD from three separate experiments. <b>F</b>, Wound healing assay was performed with MDCK cells, MDCK cells with p53-KD, or MDCK cells with mutant p53-R163H. Top panel: cell migration was determined by visual assessment of cells migrating into the wound for 24 h using a phase-contrast microscopy. Bottom panel: the time required for wound closure was measured and presented as mean ± S.D. from three separate experiments.</p

Ectopic expression of mutant p53 R261H cooperates with p53-KD to alter cyst morphology.

<p><b>A</b>, Generation of MDCK cell lines in which siRNA-resistant mutant p53 R261H was expressed along with knockdown of endogenous wild-type p53. The levels of wide-type p53 and mutant p53 R261H were determined by Western blotting. <b>B</b>, The level of wild-type p53 transcripts was determined by RT-PCR. <b>C</b>, Representative images of MDCK cells or MDCK cells with p53-KD-(R261H) in 2-D culture. <b>D</b>, Representative images of MDCK cells with p53-KD-R261H in 3-D culture for 12 d. Scale bar: 100 µM. <b>E</b>, Top panel: colony formation assay was performed with MDCK cells or MDCK cells with p53-KD-R261H. Bottom panel: the number of colonies was counted and presented as Mean ± SD from three separate experiments. <b>F</b>, Wound healing assay was performed with MDCK cells, MDCK cells with p53-KD, or MDCK cells with p53-KD-R261H. Top panel: cell migration was determined by visual assessment of cells migrating into the wound for 24 h using a phase-contrast microscopy. Bottom panel: the time required for wound closure was measured and presented as mean ± SD from three separate experiments.</p

Diverse structures, magnetism and photoluminescence of four transition metal coordination compounds based on the semirigid 4-(pyridin-3-yloxy)-phthalic acid

<div><p>Four transition metal coordination compounds, {[Co(PPDA)(H₂O)₂]}_n (<b>1</b>), {[Ni(HPPDA)₂]}_n (<b>2</b>), {[Cd(PPDA)(H₂O)]∙H₂O}_n (<b>3</b>) and {Zn(HPPDA)₂(H₂O)₄}_n (<b>4</b>), were synthesized by assembling transition metal salts with a semirigid ligand 4-(pyridin-3-yloxy)-phthalic acid (H₂PPDA) under hydrothermal conditions. The compounds have been characterized by elemental analyses, IR spectra, TGA, powder X-ray diffraction (PXRD) and single crystal X-ray crystallography. Compound <b>1</b> exhibits a 3-connected 2-D layered structure, <b>2</b> shows a (3,6)-connected 2-D layered structure, <b>3</b> displays a (3,6)-connected 2-D layered framework based on binuclear units, and <b>4</b> is a mononuclear structure, connected to generate a 3-D supramolecular architecture by hydrogen bonds. Compound <b>2</b> is thermally stable up to 300 °C. The magnetic properties of <b>1</b> and photoluminescent properties of <b>3</b> and <b>4</b> have been explored.</p></div

Additional file 2: of Complete mitochondrial genomes of eight seahorses and pipefishes (Syngnathiformes: Syngnathidae): insight into the adaptive radiation of syngnathid fishes

Statistics of the codon usage in the mitochondrial genes. (XLSX 13 kb

Arsenic trioxide cooperates with HSP90 or HDAC inhibitor to decrease mutant p53 expression and tumor cell proliferation.

<p>(A–B) Western blots were prepared with extracts from HaCaT (A) and MIA PaCa-2 (B) cells, which were untreated or treated with 1 µM 17AAG or 2 µM SAHA for 12 h. The blots were then probed with antibodies against Pirh2 and actin, respectively. (C–D) Western blots were prepared with extracts from HaCaT (C) and MIA PaCa-2 (D) cells, which were untreated or treated with 7.5 µM ATO, 1 µM 17AAG or 2 µM SAHA, alone or in combination for 12 h. The blots were then probed with antibodies against p53 and actin, respectively. (E–F) HaCaT (E) and MIA PaCa-2 (F) cells were treated as in (C–D) for 24 h. Surviving cells from both control and treated groups were counted and presented as Mean ± SD from three separate experiments. *, <i>p</i><0.05.</p

PUMA is necessary for morphogenesis of MCF10A cells.

<p><b>A</b>, Generation of MCF10A cells in which PUMA (clones #2 and 3) was stably knocked down. Western blots were performed with extracts from MCF10A cells untreated or treated with 0.2 µM doxorubicin for 24 h and then probed with antibodies against PUMA, ΔNp73 and actin, respectively. <b>B,</b> Representative images of MCF10A cells or MCF10A cells with PUMA-KD in 2-D culture (a and d, 200×) and 3-D culture (b and e, 40×; c and f, 100×). Black arrow indicates elongated spindle–liked MCF10A cells. <b>C,</b> Representative confocal images of cross-sections through the middle of acini stained with To-Pro-3 and antibody against E-cadherin in MCF10A cells with PUMA-KD. <b>D,</b> Representative confocal images of cross-sections through the middle of acini stained with To-Pro-3 and antibody against β-catenin in MCF10A cells with PUMA-KD. White arrows indicate the accumulation and translocation of β-catenin in acinus structure. <b>E</b>, Representative confocal images of cross-sections through the middle of acini stained with To-Pro-3 and antibody against laminin V in MCF10A cells with PUMA-KD. Scale bar, 20 µm.</p