Journal: ACS nano

Article Title: Pentiptycene Polymer/Single-Walled Carbon Nanotube Complexes: Applications in Benzene, Toluene, and o -Xylene Detection

doi: 10.1021/acsnano.0c02570

Figure Lengend Snippet: UV–vis–NIR absorption spectra of supernates of P1/CG100 (red), P2/CG100 (blue), P3/CG100 (purple), P4/CG100 (green), and P5/CG100 (orange). The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M11) and semiconducting (S22, S11) SWCNTs, respectively. Characteristic absorption peaks are labeled with the assigned chirality.

Article Snippet: The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M 11 ) and semiconducting (S 22 , S 11 ) SWCNTs, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 4. caption a7 Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Techniques: Labeling

Journal: ACS nano

Article Title: Pentiptycene Polymer/Single-Walled Carbon Nanotube Complexes: Applications in Benzene, Toluene, and o -Xylene Detection

doi: 10.1021/acsnano.0c02570

Figure Lengend Snippet: UV–vis–NIR absorption spectra of supernates of P4/HiPCO (blue), P4/CG100 (red), P4/SG65i (purple), and P4/CarboLex (green). The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M11) and semiconducting (S22, S11) SWCNTs, respectively.

Article Snippet: The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M 11 ) and semiconducting (S 22 , S 11 ) SWCNTs, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 4. caption a7 Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Techniques:

Journal: ACS nano

Article Title: Pentiptycene Polymer/Single-Walled Carbon Nanotube Complexes: Applications in Benzene, Toluene, and o -Xylene Detection

doi: 10.1021/acsnano.0c02570

Figure Lengend Snippet: Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Article Snippet: The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M 11 ) and semiconducting (S 22 , S 11 ) SWCNTs, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 4. caption a7 Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Techniques:

Journal: ACS nano

Article Title: Pentiptycene Polymer/Single-Walled Carbon Nanotube Complexes: Applications in Benzene, Toluene, and o -Xylene Detection

doi: 10.1021/acsnano.0c02570

Figure Lengend Snippet: Emission spectra (λex = 350 nm) of supernates of P4/CG100 (red), P4/HiPCO (blue), P4/SG65i (purple), and P4 solution (black) in chlorobenzene. The polymer concentration is 0.75 mg/mL in all cases.

Article Snippet: The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M 11 ) and semiconducting (S 22 , S 11 ) SWCNTs, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 4. caption a7 Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Techniques: Concentration Assay

Journal: ACS nano

Article Title: Pentiptycene Polymer/Single-Walled Carbon Nanotube Complexes: Applications in Benzene, Toluene, and o -Xylene Detection

doi: 10.1021/acsnano.0c02570

Figure Lengend Snippet: TEM images of drop-cast films of P1/SG65i (left), P2/SG65i (middle), and P4/SG65i (right). Inset figures are at higher magnification and show periodic fringes of SWCNTs.

Article Snippet: The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M 11 ) and semiconducting (S 22 , S 11 ) SWCNTs, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 4. caption a7 Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Techniques:

Journal: ACS nano

Article Title: Pentiptycene Polymer/Single-Walled Carbon Nanotube Complexes: Applications in Benzene, Toluene, and o -Xylene Detection

doi: 10.1021/acsnano.0c02570

Figure Lengend Snippet: Chemiresistive responses of pristine SG65i (black), P1/SG65i (red), P2/SG65i (green), and P4/SG65i (blue) in response to benzene (left), toluene (middle), and o-xylene (right) in air. Devices were exposed to analyte at 100 ppm in air for 1 min (N ≥ 6).

Article Snippet: The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M 11 ) and semiconducting (S 22 , S 11 ) SWCNTs, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 4. caption a7 Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Techniques:

Journal: ACS nano

Article Title: Pentiptycene Polymer/Single-Walled Carbon Nanotube Complexes: Applications in Benzene, Toluene, and o -Xylene Detection

doi: 10.1021/acsnano.0c02570

Figure Lengend Snippet: (a) Chemiresistive responses of P4/SG65i at different concentrations of benzene. The enlarged responses to benzene vapor at 5 ppm are shown in the inset. (b) Summary of chemiresistive responses of P1/SG65i (red), P2/SG65i (green), and P4/SG65i (blue) at different concentrations of benzene. Devices were exposed to benzene in dry air for 1 min (N ≥ 6).

Article Snippet: The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M 11 ) and semiconducting (S 22 , S 11 ) SWCNTs, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 4. caption a7 Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Techniques:

Journal: ACS nano

Article Title: Pentiptycene Polymer/Single-Walled Carbon Nanotube Complexes: Applications in Benzene, Toluene, and o -Xylene Detection

doi: 10.1021/acsnano.0c02570

Figure Lengend Snippet: Chemiresistive responses of pristine SG65i (white), P1/SG65i (blue), P2/SG65i (orange), and P4/SG65i (green) in response to different volatile organic vapors. Devices were exposed to analyte at 100 ppm in dry air for 1 min (N ≥ 6).

Article Snippet: The pink, green, and cyan boxes indicate the locations of optical transitions of metallic (M 11 ) and semiconducting (S 22 , S 11 ) SWCNTs, respectively. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 4. caption a7 Photographs of supernates of P4/HiPCO, P4/CG100, P4/SG65i, P4/CarboLex, and P4 solution in chlorobenzene (from left to right).

Techniques:

Journal: Molecular Biology of the Cell

Article Title: Inositol polyphosphate multikinase regulation of Trypanosoma brucei life stage development

doi: 10.1091/mbc.E17-08-0515

Figure Lengend Snippet: Development of BFs to PFs depends on IPMK catalytic activity. (A) Alignment of catalytic sites of IPMKs of T. brucei , A. thaliana (gene ID: AED91147.1) , Homo sapiens (gene ID: NP_689416.1), and Saccharomyces cerevisiae (gene ID: PJP09331.1). Mutated sites are shown in red. (B) Recombinant T. brucei IPMK WT or mutants (37 kDa) resolved on a 4–20% SDS–PAGE gel stained with Coomassie Imperial Staining (Pierce). (C) Activity analysis of rIPMK WT and mutants with Ins(1,4,5)P3 and Ins(1,3,4,5)P4 substrates. See for kinetic parameters of each rIPMK enzyme and substrates. Growth curve (D) and Western blot analysis (E) of T. brucei IPMK CN exclusively expressing V5-tagged IPMK WT, mutants D142A, K164W, D142A/K144A, or no IPMK gene (knockdown). Cells were grown in the absence of tet for exclusive expression of introduced V5-tagged WT, mutant allele, or IPMK knockdown. Blots were stripped and reblotted with mAb 78, which recognizes mitochondrial HSP70. (F) Flow cytometry analysis and quantification of T. brucei expressing EP procyclin after IPMK knockdown or exclusive expression (EE) of WT or mutant IPMK alleles for 36 h in HMI-9 at 37°C in BF followed by 6 d in SDM-79 at 27°C.

Article Snippet: For confirmation of IP–protein binding, lysates of 5.0 × 10 7 mid–log growth T. brucei BFs were incubated with biotin-Ins(1,4,5)P3 or biotin-Ins(1,3,4,5)P4 (Echelon Biosciences; biotin linked through the 1-position on the inositol ring) in binding buffer (as above) followed by incubation with 50 µl of Dynabeads M-270 Streptavidin (ThermoFisher Scientific).

Techniques: Activity Assay, Recombinant, SDS Page, Staining, Western Blot, Expressing, Mutagenesis, Flow Cytometry

Journal: Molecular Biology of the Cell

Article Title: Inositol polyphosphate multikinase regulation of Trypanosoma brucei life stage development

doi: 10.1091/mbc.E17-08-0515

Figure Lengend Snippet: Proteomic analysis of inositol phosphate interacting proteins. (A) Volcano plots indicate proteins that interact with Ins(1,4,5)P3 or Ins(1,3,4,5)P4 compared with nonconjugated beads (enrichment cutoff ≥ 2-fold and p value < 0.1). See Supplemental Dataset S2 for list of proteins. (B) Fold enrichment analysis of proteins that interact with each IP by functional categories. (C) Network analysis of protein IP interactions. Blue line, Ins(1,3,4,5)P4 interactions; red line, Ins(1,3,4)P3 interaction; gray line, proteins that interact with both IPs. (D) Ratio of fold enrichment of proteins identified as interacting with Ins(1,3,4,5)P4 vs. Ins(1,4,5)P3. (E) GO enrichment analysis of proteins interacting with Ins(1,4,5)P3, Ins(1,3,4,5)P4, or both. (F) Heat map of metabolic proteins that interact with each IP (subset indicated by brackets in B). Fold enrichment values are indicated for each protein.

Article Snippet: For confirmation of IP–protein binding, lysates of 5.0 × 10 7 mid–log growth T. brucei BFs were incubated with biotin-Ins(1,4,5)P3 or biotin-Ins(1,3,4,5)P4 (Echelon Biosciences; biotin linked through the 1-position on the inositol ring) in binding buffer (as above) followed by incubation with 50 µl of Dynabeads M-270 Streptavidin (ThermoFisher Scientific).

Techniques: Functional Assay

Journal: Journal of Virology

Article Title: Murine Cytomegalovirus M25 Proteins Sequester the Tumor Suppressor Protein p53 in Nuclear Accumulations

doi: 10.1128/JVI.00574-20

Figure Lengend Snippet: M25 proteins interact with p53. (A) NIH 3T3 cells were infected with the indicated viruses (at MOI 1) and harvested 36 h p.i. GFP and GFP-tagged M25 proteins were precipitated with GFP nanobodies, and immunoblotting was performed with antibodies for the specified proteins. (B) p53-deficient HCT116 cells were transfected with plasmids coding for the indicated proteins, and immunoprecipitation was performed with an HA (top panel) or a myc antibody (bottom panel) using cell lysates prepared 24 h posttransfection. Antibodies used for immunoblotting are indicated in the right margin. IP, precipitated proteins; IN, input lysates. (C) MEF were infected with WT MCMV or the ΔM25 mutant at an MOI of 1 or were left uninfected, and at 36 h p.i. the cells were labeled with the indicated antibodies and analyzed by confocal microscopy. Images to the left are depicted in inverse mode, with positive staining appearing in black and gray. Infected cells were identified based on GFP expression (right panels). Please note that the apparently different GFP distribution is due to the different effects of WT MCMV and the ΔM25 mutant on cell morphology. While WT MCMV-infected cells become spherical, leaving a small cytoplasmic rim around the cell nucleus, ΔM25-infected cells retain an elongated shape. (D) NIH 3T3 cells were infected with the tagged virus vM25GFP for 24 h, followed by immunolabeling with an antibody against the E1 protein. The M25-GFP fusion protein was detected via GFP fluorescence. Shapes of the nuclei were determined based on Hoechst staining. Images are representative of at least 20 cells analyzed per condition. Scale bars, 10 μm.

Article Snippet: Plasmid pCMV6-Entry-Trp53-Myc-DKK coding for murine p53 was purchased from OriGene (MR206086).

Techniques: Infection, Western Blot, Transfection, Immunoprecipitation, Mutagenesis, Labeling, Confocal Microscopy, Staining, Expressing, Immunolabeling, Fluorescence

Journal: Journal of Virology

Article Title: Murine Cytomegalovirus M25 Proteins Sequester the Tumor Suppressor Protein p53 in Nuclear Accumulations

doi: 10.1128/JVI.00574-20

Figure Lengend Snippet: p53 accumulates in WT MCMV-infected cells and in M25-expressing cell lines. (A and B) NIH 3T3 cells were infected with WT MCMV or the ΔM25 mutant (MOI of 3) for the indicated time periods, and lysates were analyzed by immunoblotting with the indicated antibodies. GAPDH served as a loading control, and the viral M25 and M55 proteins were used as infection marker. (C) Using the indicated viruses NIH 3T3 cells were infected as in panels A and B, and cell lysates prepared at 36 h p.i. were analyzed by immunoblotting with antibodies specific for the indicated proteins. The viral E1 protein served as an infection marker. (D) NIH 3T3-derived cell lines encoding pM25l-HA, pM25s-HA, or GFP and parental NIH 3T3 cells were treated with doxycycline (Doxy) for 24 h or left untreated as indicated. Doxorubicin (Doxr) treatment of NIH 3T3 cells (for 24 h) was used for inducing genotoxic stress (positive control). Cell lysates were analyzed by immunoblotting using the indicated antibodies. The HA antibody detected the M25 proteins, and ERK1/2 (extracellular signal-regulated kinase 1/2) served as a loading control. (E) The cell lines expressing the proteins indicated at the top and NIH 3T3 cells (panels at the right) were treated with doxycycline for 24 h, followed by immunolabeling with the indicated antibodies. GFP was detected using its autofluorescence, and doxorubicin (Doxr) treatment of NIH 3T3 cells was used as a positive control to induce p53 expression (second last panels on the right). Scale bars, 10 μm.

Article Snippet: Plasmid pCMV6-Entry-Trp53-Myc-DKK coding for murine p53 was purchased from OriGene (MR206086).

Techniques: Infection, Expressing, Mutagenesis, Western Blot, Marker, Derivative Assay, Positive Control, Immunolabeling

Journal: Journal of Virology

Article Title: Murine Cytomegalovirus M25 Proteins Sequester the Tumor Suppressor Protein p53 in Nuclear Accumulations

doi: 10.1128/JVI.00574-20

Figure Lengend Snippet: The half-life of p53 is increased in WT MCMV-infected cells and in M25-expressing cells. (A) NIH 3T3 cells were used either uninfected or were infected with WT MCMV or ΔM25 MCMV. At 18 h p.i., the cells were cultivated for another 6 h or treated with Nutlin3a (40 μM), followed by immunoblot analysis of cell lysates with the indicated antibodies. Vinculin served as a loading control, and the viral E1 early protein served as an infection marker. (B) NIH 3T3 cells either uninfected or infected as described previously (see panel A) for 24 h were then treated with cycloheximide (CHX; t = 0 h). At the specified time points thereafter, lysates of cells were prepared and analyzed by immunoblotting with the indicated antibodies. GAPDH served as a loading control, and the viral immediate early protein IE1 was used as an infection marker. (C) Parental NIH 3T3 cells and cell lines encoding the indicated proteins were treated with doxycycline for 24 h, followed by treatment with CHX for the indicated time periods (min) and analysis of p53 amounts by immunoblotting. ERK1/2 served as a loading control. The noninduced sample remained untreated (without doxycycline) and consisted of a mixture of cells transduced with M25l-, M25s-, or GFP-encoding lentiviral vectors (one third each). Doxorubicin (Doxr) treatment of NIH 3T3 cells was used for activation of p53 (bottom row). (A to C) The blots are representative of 3 independently performed experiments. (B and C) Signals were quantified by densiometric analysis using ImageJ and normalized to the respective loading control. p53 levels are depicted in relation to the amounts present at the time point when CHX was added (time point 0), and values represent the means of values from three independent experiments.

Article Snippet: Plasmid pCMV6-Entry-Trp53-Myc-DKK coding for murine p53 was purchased from OriGene (MR206086).

Techniques: Infection, Expressing, Western Blot, Marker, Transduction, Activation Assay

Journal: Journal of Virology

Article Title: Murine Cytomegalovirus M25 Proteins Sequester the Tumor Suppressor Protein p53 in Nuclear Accumulations

doi: 10.1128/JVI.00574-20

Figure Lengend Snippet: Influence of M25 proteins on posttranslational modification of p53 and its transcriptional activity. (A) NIH 3T3 cells were either left uninfected or were infected with WT MCMV or the ΔM25 mutant (MOI of 3) or were treated with doxorubicin (Doxr; 0.5 μM for 24 h) or with Nutlin3a (Nut3a; 40 μM for 6 h). Cell lysates prepared at the indicated time points postinfection were analyzed by immunoblotting with antibodies for the specified proteins and the modified p53 versions. (B) p53-deficient HCT116 cells were transfected with the luciferase reporter plasmid PG13-luc, a Gaussia luciferase control plasmid and expression plasmids encoding p53, pM25s, pM25l, or the adenovirus E1B-55K protein as indicated. The luciferase activities were measured 48 h after transfection. The graph shows luciferase activities ± the standard deviations (SD) in cell lysates (ratios of firefly and Gaussia luciferase activity) of three biological replicates. A Mann-Whitney test (one-tailed) was used to analyze statistical significance. (C) Total RNA was isolated from NIH 3T3 cells infected with WT MCMV or ΔM25 (MOI of 3) for 36 h and, for comparison, from uninfected cells (either untreated or treated with Nutlin3a [5 μM]) and was subjected to qRT PCR analysis using gene-specific primers. (D) Total RNA was isolated from primary MEF infected with either WT MCMV or ΔM25 (MOI of 3) for the depicted duration and from cells that remained uninfected or were treated with doxorubicin (Doxr) for 24 h. qRT PCR was performed using Cdkn1a-specific primers. (C and D) The graphs depict the ΔΔCT values of the qRT-PCRs (log2-transformed values of expression levels in relation to the ones of uninfected cells). The means ± the SD of three to five biological replicates are shown. A t test (two-tailed) was used for statistical analysis. (E) NIH 3T3 cells were infected or treated as indicated in panel A, and cell lysates prepared at the indicated time points postinfection were analyzed by immunoblotting with antibodies specific for PUMA or BAX.

Article Snippet: Plasmid pCMV6-Entry-Trp53-Myc-DKK coding for murine p53 was purchased from OriGene (MR206086).

Techniques: Modification, Activity Assay, Infection, Mutagenesis, Western Blot, Transfection, Luciferase, Plasmid Preparation, Expressing, MANN-WHITNEY, One-tailed Test, Isolation, Quantitative RT-PCR, Transformation Assay, Two Tailed Test

Journal: Journal of Virology

Article Title: Murine Cytomegalovirus M25 Proteins Sequester the Tumor Suppressor Protein p53 in Nuclear Accumulations

doi: 10.1128/JVI.00574-20

Figure Lengend Snippet: Effect of M25 proteins on p53 association with promoter sequences. NIH 3T3 were either used uninfected or were infected with WT MCMV or the ΔM25 mutant (MOI of 3) or were treated with doxorubicin (Doxr) for 18 h. Chromatin immunoprecipitation (ChIP) was performed using a p53 antibody or IgG for control, and the amounts of precipitated and input DNA were quantified by qPCR. The values represent the percentage of precipitated DNA (means plus the SD of four biological replicates). A t test (one-tailed) was used for statistical analysis.

Article Snippet: Plasmid pCMV6-Entry-Trp53-Myc-DKK coding for murine p53 was purchased from OriGene (MR206086).

Techniques: Infection, Mutagenesis, Chromatin Immunoprecipitation, One-tailed Test

Journal: Journal of Virology

Article Title: Murine Cytomegalovirus M25 Proteins Sequester the Tumor Suppressor Protein p53 in Nuclear Accumulations

doi: 10.1128/JVI.00574-20

Figure Lengend Snippet: Growth of the ΔM25 mutant and of WT MCMV on p53-deficient MEF. p53-negative (p53–/–) and p53-positive (p53+/–) MEF were infected with the indicated viruses (MOI of 1), and titers of infectious virus in the supernatant of cultures at the indicated time points postinfection were determined by plaque assay. Average titers (means) ± the SD of triplicate cultures are depicted. The data are from one experiment representative of three independent experiments performed.

Article Snippet: Plasmid pCMV6-Entry-Trp53-Myc-DKK coding for murine p53 was purchased from OriGene (MR206086).

Techniques: Mutagenesis, Infection, Plaque Assay