Activation of the p21 promoter by JMJD2D in HEK293T cells.

<p>(<b>A</b>) Activity of a p21 luciferase reporter construct upon cotransfection of control vector pEV3S or wild-type or H192A JMJD2D is depicted. As indicated, empty vector pcDNA3 or pcDNA3-p53 was also transfected. (<b>B</b>) Analogous, response of a CMV or MMP-1 luciferase reporter construct to cotransfection of p53 and/or JMJD2D. (<b>C</b>) Chromatin immunoprecipitation assay in HCT116 cells treated without and with adriamycin.</p

Mapping of interaction domains.

<p>(<b>A</b>) Indicated Flag-tagged amino acids of JMJD2D were coexpressed with p53 in HEK293T cells and complex formation assessed in coimmunoprecipitation assays as in Fig. 1B. A sketch of human JMJD2D highlighting its JmjC domain is presented at the bottom. (<b>B</b>) Comparable amounts of GST or indicated GST-p53 fusion proteins were bound to glutathione agarose. After incubation with Flag-tagged JMJD2D, bound JMJD2D was revealed by anti-Flag Western blotting. The location of the DNA binding domain within p53 is indicated in the sketch at the bottom.</p

JMJD2D depletion results in reduced cell proliferation.

<p>(<b>A</b>) HCT116 cells expressing control or JMJD2D shRNA were treated with 1 µM adriamycin or with DMSO as a control for 24 h. Downregulation of JMJD2D was assessed by Western blotting; immunoblotting for actin served as a loading control. (<b>B</b>) Wild-type or p53^−/− HCT116 cells were challenged with control or JMJD2D shRNA (#1 or #3) and then treated without or with 1 µM adriamycin for 72 h. The number of cells were counted and presented as percent of the control shRNA for each wild-type and p53^−/− HCT116 cells. Statistical significance of differences between various experimental conditions is indicated in the graph.</p

JMJD2D protects from apoptosis.

<p>(<b>A</b>) The level of sub-G1 HCT116 cells was determined in cells that expressed control shRNA or JMJD2D shRNA#3. Cells were treated for 72 h with 1 µM adriamycin or DMSO as indicated. (<b>B</b>) The same in case of p53^−/− HCT116 cells. (<b>C</b>) Model of JMJD2D action.</p

Stimulation of p21 expression by JMJD2D in U2OS cells.

<p>(<b>A</b>) Wild-type or H192A JMJD2D or empty vector pEV3S were stably transfected into U2OS cells. Expression of indicated proteins was assessed by Western blotting in cells treated for 24 h with 1 µM adriamycin or DMSO as a control. (<b>B</b>) RNA was isolated from stably transfected U2OS cells and RT-PCR analyses were performed. Shown is the amplification of p21 cDNA and, as a control, GAPDH cDNA.</p

Binding of JMJD2D to p53.

<p>(<b>A</b>) Flag-tagged JMJD2D or HSPBAP1 were coexpressed with p53 in HEK293T cells. After anti-p53 immunoprecipitation (IP), coprecipitated proteins were revealed by anti-Flag immunoblotting (top panel). The bottom two panels show input levels of Flag-tagged proteins or p53. IgH, immunoglobulin heavy chain. (<b>B</b>) Indicated Flag-tagged JMJD proteins were coexpressed with p53 in HEK293T cells. After anti-Flag immunoprecipitation, coprecipitated p53 was detected by anti-p53 Western blotting (top panel). The middle and bottom panels show p53 and Flag-JMJD protein input levels, respectively. (<b>C</b>) HCT116 cell extracts were challenged with no, control or anti-JMJD2D antibodies and coprecipitated p53 detected by immunoblotting (top panel). The bottom panel shows that respective JMJD2D input levels were equal.</p

DataSheet_1_Methylation of the epigenetic JMJD2D protein by SET7/9 promotes prostate tumorigenesis.pdf

How the function of the JMJD2D epigenetic regulator is regulated or whether it plays a role in prostate cancer has remained elusive. We found that JMJD2D was overexpressed in prostate tumors, stimulated prostate cancer cell growth and became methylated by SET7/9 on K427. Mutation of this lysine residue in JMJD2D reduced the ability of DU145 prostate cancer cells to grow, invade and form tumors and elicited extensive transcriptomic changes. This included downregulation of CBLC, a ubiquitin ligase gene with hitherto unknown functions in prostate cancer, and upregulation of PLAGL1, a transcription factor with reported tumor suppressive characteristics in the prostate. Bioinformatic analyses indicated that CBLC expression was elevated in prostate tumors. Further, downregulation of CBLC largely phenocopied the effects of the K427 mutation on DU145 cells. In sum, these data have unveiled a novel mode of regulation of JMJD2D through lysine methylation, illustrated how this can affect oncogenic properties by influencing expression of the CBLC gene, and established a pro-tumorigenic role for CBLC in the prostate. A corollary is that JMJD2D and CBLC inhibitors could have therapeutic benefits in the treatment of prostate and possibly other cancers.</p

Phenyltetraene-Based Nonlinear Optical Chromophores with Enhanced Chemical Stability and Electrooptic Activity

“Push−pull” phenyltetraene-based chromophores are too sensitive to be incorporated into Diels−Alder-type cross-linkable polymers due to the reactivity of its diene segment with maleimides. A facile synthetic route has been explored to incorporate a methoxy group into the R position of such chromophores, which reduces their diene reactivity during the poling and lattice hardening process. The poled polymers with one of such chromophores doped in a cross-linked polymer lattice showed ultrahigh electro-optic activities, up to 306 pm/V at 1310 nm

Diels−Alder “Click Chemistry” for Highly Efficient Electrooptic Polymers

Diels−Alder “Click Chemistry” for Highly Efficient Electrooptic Polymer

Ultrafast Exciton Dissociation Followed by Nongeminate Charge Recombination in PCDTBT:PCBM Photovoltaic Blends

The precise mechanism and dynamics of charge generation and recombination in bulk heterojunction polymer:fullerene blend films typically used in organic photovoltaic devices have been intensively studied by many research groups, but nonetheless remain debated. In particular the role of interfacial charge-transfer (CT) states in the generation of free charge carriers, an important step for the understanding of device function, is still under active discussion. In this article we present direct optical probes of the exciton dynamics in pristine films of a prototypic polycarbazole-based photovoltaic donor polymer, namely poly[N-11′′-henicosanyl-2,7-carbazole-alt-5,5-(4′,7′-di-2-thienyl-2′,1′,3′-benzothiadiazole)] (PCDTBT), as well as the charge generation and recombination dynamics in as-cast and annealed photovoltaic blend films using methanofullerene (PC61BM) as electron acceptor. In contrast to earlier studies we use broadband (500–1100 nm) transient absorption spectroscopy including the previously unobserved but very important time range between 2 ns and 1 ms, which allows us not only to observe the entire charge carrier recombination dynamics but also to quantify the existing decay channels. We determine that ultrafast exciton dissociation occurs in blends and leads to two separate pools of products, namely Coulombically bound charge-transfer (CT) states and unbound (free) charge carriers. The recombination dynamics are analyzed within the framework of a previously reported model for poly(3-hexylthiophene):PCBM (Howard, I. A. J. Am. Chem. Soc. 2010, 132, 14866) based on concomitant geminate recombination of CT states and nongeminate recombination of free charge carriers. The results reveal that only ∼11% of the initial photoexcitations generate interfacial CT states that recombine exclusively by fast nanosecond geminate recombination and thus do not contribute to the photocurrent, whereas ∼89% of excitons create free charge carriers on an ultrafast time scale that then contribute to the extracted photocurrent. Despite the high yield of free charges the power conversion efficiency of devices remains moderate at about 3.0%. This is largely a consequence of the low fill factor of devices. We relate the low fill factor to significant energetic disorder present in the pristine polymer and in the polymer:fullerene blends. In the former we observed a significant spectral relaxation of exciton emission (fluorescence) and in the latter of the polaron-induced ground-state bleaching, implying that the density of states (DOS) for both excitons and charge carriers is significantly broadened by energetic disorder in pristine PCDTBT and in its blend with PCBM. This disorder leads to charge trapping in solar cells, which in turn causes higher carrier concentrations and more significant nongeminate recombination. The nongeminate recombination has a significant impact on the IV curves of devices, namely its competition with charge carrier extraction causes a stronger bias dependence of the photocurrent of devices, in turn leading to the poor device fill factor. In addition our results demonstrate the importance of ultrafast free carrier generation and suppression of interfacial CT-state formation and question the applicability of the often used Braun–Onsager model to describe the bias dependence of the photocurrent in polymer:fullerene organic photovoltaic devices