Journal: Cell Death & Disease

Article Title: A novel angiogenesis inhibitor impairs lovo cell survival via targeting against human VEGFR and its signaling pathway of phosphorylation

doi: 10.1038/cddis.2012.145

Figure Lengend Snippet: HMQ18–22 inhibited cell viability and decreased phosphorylation of VEGFR2, VEGFR1, Akt, PKC α and PLC γ -1 involved in angiogenesis. ( a ) HMQ18–22 decreased cell survival in lovo and HUVEC cells. ( b ) The AlphaScreen signal indicated HMQ18–22 decreased VEGFR phosphorylation. ( c ) Cell were treated with VEGF (50 ng/ml) for 30 min before extracting proteins with RIPA lysis buffer. HMQ18–22 decreased the phosphorylation of VEGFR2(Tyr 1214 ), VEGFR1(Tyr 1333 ), Akt(Tyr 326 ), PKC α (Tyr 657 ) and PLC γ -1(Tyr 771 ) by western blot analysis. On the contrary, the Raf1(Tyr 341 ) phosphorylation was not altered by HMQ18–22. Results were quantified by densitometry analysis of the bands form and then normalization to GAPDH protein. ( d ) Effect of HMQ18–22 on cells transfected with siRNAs targeting of VEGFR2, VEGFR1, Akt, PKC α or PLC γ -1. Quantification of RT-PCR data showed knockdown of VEGFR2, VEGFR1, Akt, PKC α and PLC γ -1; the bottom right panel showed the effect of HMQ18–22 on cells proliferation was attenuated in knockdown cells. Data were expressed as mean values±S.D. ( n =3). * P <0.05, ** P <0.01 versus the untreated control group.

Article Snippet: VEGFR2 kinase was from Carna Biosciences (Kobe, Japan).

Techniques: Amplified Luminescent Proximity Homogenous Assay, Lysis, Western Blot, Transfection, Reverse Transcription Polymerase Chain Reaction

Journal: Cell Death & Disease

Article Title: A novel angiogenesis inhibitor impairs lovo cell survival via targeting against human VEGFR and its signaling pathway of phosphorylation

doi: 10.1038/cddis.2012.145

Figure Lengend Snippet: HMQ18–22 inhibited tumor growth in nude mice bearing human colon cancer xenografts. ( a ) The representative xenografts of lovo human colon cancer in mice. ( b ) HMQ18–22 decreased the phosphorylation of VEGFR2(Tyr 1214 ), VEGFR1(Tyr 1333 ), Akt(Tyr 326 ), PKC α (Tyr 657 ) and PLC γ -1(Tyr 771 ) in the tumor tissues by western blot analysis. ( c ) Quantitation data of ( b ). Data were expressed as mean values±S.D. ( n =3). ** P <0.01 versus the untreated control

Article Snippet: VEGFR2 kinase was from Carna Biosciences (Kobe, Japan).

Techniques: Western Blot, Quantitation Assay

Journal: Weed Research

Article Title: Interpopulation variability and adaptive potential for reduced glyphosate sensitivity in Alopecurus myosuroides

doi: 10.1111/wre.12264

Figure Lengend Snippet: Collection sites of 40 UK Alopecurus myosuroides populations collected in July 2012 and tested for glyphosate sensitivity using dose–response analysis.

Article Snippet: Glyphosate [Roundup ProBiactive, 360 g L −1 glyphosate present as the isopropylamine salt at 480 g L −1 (41.1% w/w) (Monsanto)] was applied with an experimental track sprayer (generation III research sprayer, DeVries) using a flat fan nozzle (FE80/0.8/3) calibrated to deliver 200 L ha −1 at a pressure of 300 KPa.

Techniques:

Journal: Weed Research

Article Title: Interpopulation variability and adaptive potential for reduced glyphosate sensitivity in Alopecurus myosuroides

doi: 10.1111/wre.12264

Figure Lengend Snippet: Dose–response curves (Weibull 2‐parameter function) for six Alopecurus myosuroides populations subjected to recurrent glyphosate selection (T1, one generation of selection, T2, two generations of selection, C2, control, unselected line). Symbols represent mean observed survival data and lines are fitted regression models. (A) Population ACA212; (B) Population ACA412; (C) Population AES112; (D) Population AES212; (E) Population ASF112; (F) Population AWA712.

Article Snippet: Glyphosate [Roundup ProBiactive, 360 g L −1 glyphosate present as the isopropylamine salt at 480 g L −1 (41.1% w/w) (Monsanto)] was applied with an experimental track sprayer (generation III research sprayer, DeVries) using a flat fan nozzle (FE80/0.8/3) calibrated to deliver 200 L ha −1 at a pressure of 300 KPa.

Techniques: Selection, Control

Journal: Weed Research

Article Title: Interpopulation variability and adaptive potential for reduced glyphosate sensitivity in Alopecurus myosuroides

doi: 10.1111/wre.12264

Figure Lengend Snippet: Estimated ED 50 values for glyphosate‐selected and control Alopecurus myosuroides lines. Error bars are standard errors. T‐tests were performed to compare ED 50 estimates for the control (C2) populations to the once‐selected (T1) and twice‐selected (T2) lines (** P < 0.01, *** P < 0.001).

Article Snippet: Glyphosate [Roundup ProBiactive, 360 g L −1 glyphosate present as the isopropylamine salt at 480 g L −1 (41.1% w/w) (Monsanto)] was applied with an experimental track sprayer (generation III research sprayer, DeVries) using a flat fan nozzle (FE80/0.8/3) calibrated to deliver 200 L ha −1 at a pressure of 300 KPa.

Techniques: Control

Journal: Cells

Article Title: Identification of Novel Positive Allosteric Modulators of Neurotrophin Receptors for the Treatment of Cognitive Dysfunction

doi: 10.3390/cells10081871

Figure Lengend Snippet: Mechanism of action of triazinetriones on TrkA. ( a ) Full-length TrkA with a HA-tag (TrkA-HA) fused to the C -terminus was purified through immunoprecipitation using anti-HA agarose beads. Purified TrkA-HA was incubated with DMSO (blue circles and hatched line), ACD855 (5 µM) (black triangles and solid line), or ACD856 (1 µM) (red squares and solid line) for approx. 5 min. Thereafter, ATP was added to yield the indicated concentration. Each data point is the mean ± SEM ( n = 3). The solid lines are the curve-fit using the Michaelis–Menten equation used to calculate the apparent Vmax(app) and km(app), and the dotted lines are the 95% confidence band for each curve fit. ( b – d ) Affinity labeling and streptavidin adsorption of Trka. ( b ) Western blot of streptavidin adsorbed TrkA-HA non-covalent labeled with NHS-biotinylated triazinetrione compound (lane 2) or covalent labeled by UV-crosslinking of 100 µM sulfo-SBED biotinylated compound (lane 3). Detection of immunoreactive band was performed with anti-TrkA antibody. Supernatant loaded to the left (lane 1) was used as positive control for the Western blot. Arrows on the left indicate the migration of molecular weight markers corresponding to 198, 98, and 62 kDa. ( c ) Anti-HA agarose immunoprecipitation of cross-linked or non-crosslinked sulfo-SBED compound (AC27019-SBED) from cell lysate incubated with 100 µM AC27019-SBED. Lane 1, non-UV-crosslinking, and lane 2, UV-crosslinking of sulfo-SBED labeled TrkA-HA, both detected with streptavidin-HRP. Lanes 3 and 4 are loading controls of lanes 1 and 2, respectively, were TrkA-HA was detected by immunoblotting using the anti-TrkA antibody. Both blots were part of the same gel, but the membrane was cut in two pieces, and the proteins were detected by streptavidin-HRP (left panel) or by an anti-TrkA antibody (right panel). ( d ) Streptavidin adsorption of biotinylated compound bound to TrkA-HA. Lane 1, cell lysate used for positive control of immunodetection; lane 3, cell lysate without biotinylated compound was adsorbed to streptavidin-agarose as negative control; lanes 5 and 7, cell lysates containing sulfo-SBED compound UV-crosslinked to TrkA (from two different experiments) were adsorbed to streptavidin-agarose and immunoblotted using anti-TrkA antibody. Lanes 2, 4, and 6 are empty lanes to avoid cross-contamination between lanes.

Article Snippet: Recombinant intracellular domain (ICD) of TrkA (08-186) and single site biotinylated activated (08-486-20N) or non-activated TrkA (08-486-23N) were obtained from Carna Biosciences, Odense, Denmark.

Techniques: Purification, Immunoprecipitation, Incubation, Concentration Assay, Labeling, Adsorption, Western Blot, Positive Control, Migration, Molecular Weight, Membrane, Immunodetection, Negative Control