Journal: Scientific Reports

Article Title: Malaria risk assessment and mapping using satellite imagery and boosted regression trees in the Peruvian Amazon

doi: 10.1038/s41598-019-51564-4

Figure Lengend Snippet: Predictor and outcome variables used in BRT models.

Article Snippet: Shortest distance to rivers (SDR) , environmental variable estimated using the Global Surface Water (occurrence) map from JRC (Joint Research Centre).

Techniques:

Journal: Human molecular genetics

Article Title: The huntingtin interacting protein HIP1 is a clathrin and alpha-adaptin-binding protein involved in receptor-mediated endocytosis.

doi: 10.1093/hmg/10.17.1807

Figure Lengend Snippet: Figure 2. Affinity purification and identification of HIP1-binding proteins. (A) Affinity purification of GST-HIP1 (218–604) interacting proteins. Human brain extract was incubated with 5 (lane 1), 25 (lane 2) or 50 µg (lane 3) GST-HIP1 protein immobilized on 25 µl packed glutathione-agarose. After washing of the beads, bound proteins were eluted with SDS-sample buffer and analyzed by SDS–PAGE and Coomassie blue staining. As controls, the extract was incubated with glutathione–agarose beads without any immobilized protein (lane 4) or with 50 µg immobilized GST (lane 5). (B) Identification of HIP1 bound proteins by MALDI-MS. The peptide mass map of the prominent 170 kDa protein band after ‘in-gel’ digestion with trypsin is shown. Peptides labeled with asterisks matched calculated tryptic peptide masses of human clathrin heavy chain 1. (C) Identification of HIP1 bound proteins by immuno- blotting. Portions of the protein samples analyzed in (A) were separated by SDS–PAGE, blotted onto nitrocellulose and probed with anti-clathrin heavy chain, anti-α-adaptin, anti-HAP1 and anti-huntingtin(CAG53b) antibodies.

Article Snippet: Tryptic peptide mass maps of the proteins were recorded on a Bruker Scout MTP Reflex III MALDI mass spectrometer (Bruker Daltonik, Germany) using the matrix α-cyano-4hydroxycinamic acid.

Techniques: Affinity Purification, Binding Assay, Incubation, SDS Page, Staining, Labeling

Journal: Cancers

Article Title: p38 MAPK Inhibition Mitigates Hypoxia-Induced AR Signaling in Castration-Resistant Prostate Cancer

doi: 10.3390/cancers13040831

Figure Lengend Snippet: p38 MAPK inhibition decreases cell proliferation and survival in prostate cancer cells. ( a ) Western blots from DU145, PC3, VCaP, LNCaP, V16D and MR49F cells showing AR expression. ( b ) Indicated cell lines were treated with 10 µM SB203580 (SB) under normoxia (21% O 2 ) (left) or hypoxia (0.2% O 2 ) (right) for 150 h. Confluency was measured with the IncuCyte Live Cell Imaging system after confirming proportionality to cell numbers. Each data point represents an independent experiment, bars represent the mean value ± S.E.M. ( c ) V16D cells were transfected with siRNA targeting MAPK11, MAPK14, or non-targeting negative control (NTC). Expression of target genes was assessed by qPCR (left) or western blotting (right). ( d ) Cells as in (c) were placed in normoxia (21% O 2 ) or hypoxia (0.2% O 2 ) for 72 h and confluency was measured with the IncuCyte Live Cell Imaging system. ( e ) V16D cells were treated with various doses of SB203580 (5, 10, 20 µM) for 72 h under normoxia (21% O 2 ) or hypoxia (0.2% O 2 ). Single cells were seeded for clonogenic survival in triplicate and surviving fraction calculated from colony formation 14 days later. Data points represent independent experiments and bars represent the mean value ± S.E.M. (*: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001).

Article Snippet: The membrane was incubated with antibodies diluted in Odyssey blocking buffer (LI-COR Biosciences) overnight at 4 °C to detect the following proteins: p-p38 MAPK (Tyr182; E-1; 1:500; Santa Cruz Biotechnology, Dallas, TX, USA), p38 MAPK (1:1000), p-Hsp27 (Ser82; 1:1000), Hsp27 (G31; 1:1000), AR (D6F11; 1:1000), HIF-1ɑ (1:1000; BD Biosciences, San Jose, CA, USA), eIF4E (1:1000; BD Biosciences), and beta-tubulin (1:10,000; Abcam, Cambridge, UK).

Techniques: Inhibition, Western Blot, Expressing, Live Cell Imaging, Transfection, Negative Control

Journal: Cancers

Article Title: p38 MAPK Inhibition Mitigates Hypoxia-Induced AR Signaling in Castration-Resistant Prostate Cancer

doi: 10.3390/cancers13040831

Figure Lengend Snippet: p38 MAPK inhibition decreases Hsp27 phosphorylation, AR activity and expression of AR target genes under normoxia and hypoxia. ( a ) Prostate cancer cell lines (DU145, PC3, VCaP, LNCaP, V16D, MR49F) were treated with 10 µM SB203580 for one hour and subjected to western blotting (left). V16D cells were treated with 25 µM of anisomycin for one hour as a positive control for p38 MAPK activation. Densitometry analysis was performed to calculate fold change of p-Hsp27 (Ser82) relative to Hsp27 for each cell line. ( b ) V16D cells were transfected with a plasmid encoding luciferase driven by the AR promoter. 24 h later, cells were treated with 1 nM DHT, 10 µM SB203580, or 10 µM enzalutamide (Enza) in 10% charcoal treated (CT)-FBS media (androgen depleted) and incubated under normoxia (21% O 2 ) or hypoxia (0.2% O 2 ) for 48 h. Relative light units (RLU) from luciferase was measured after addition of luciferin. ( c–e ) V16D cells were treated with 10 µM SB203580, or 10 µM enzalutamide for 48 h and relative mRNA expression of KLK3 ( c ), NKx3.1 ( d ), and FKBP5 ( e ) was measured by RT-qPCR normalized to the average of HPRT1 and GUSB gene expression. Fold change was calculated relative to the 10% CT-FBS negative normoxia control. Data points represent an independent experiment and bars represent the mean value ± S.E.M. (*: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001).

Article Snippet: The membrane was incubated with antibodies diluted in Odyssey blocking buffer (LI-COR Biosciences) overnight at 4 °C to detect the following proteins: p-p38 MAPK (Tyr182; E-1; 1:500; Santa Cruz Biotechnology, Dallas, TX, USA), p38 MAPK (1:1000), p-Hsp27 (Ser82; 1:1000), Hsp27 (G31; 1:1000), AR (D6F11; 1:1000), HIF-1ɑ (1:1000; BD Biosciences, San Jose, CA, USA), eIF4E (1:1000; BD Biosciences), and beta-tubulin (1:10,000; Abcam, Cambridge, UK).

Techniques: Inhibition, Phospho-proteomics, Activity Assay, Expressing, Western Blot, Positive Control, Activation Assay, Transfection, Plasmid Preparation, Luciferase, Incubation, Quantitative RT-PCR, Gene Expression, Control

Journal: Cancers

Article Title: p38 MAPK Inhibition Mitigates Hypoxia-Induced AR Signaling in Castration-Resistant Prostate Cancer

doi: 10.3390/cancers13040831

Figure Lengend Snippet: Androgen and hypoxia activate p38 MAPK and Hsp27. ( a ) V16D cells were exposed to 0 nM, 1 nM, or 10 nM DHT in charcoal-stripped media for 6 h in normoxia (21% O 2 ) or hypoxia (0.2% O 2 ). 25 µM of anisomycin for one hour was used as a positive control for p38 MAPK activation. Total cell lysates were subjected to western blotting. Densitometry analysis of protein bands was performed to calculate fold change of p-p38 (Tyr182) ( b ) and p-Hsp27 (Ser82) ( c ) relative to normoxia control with 0 nM DHT and normalized to eIF4E. Data points represent independent experiments and bars represent the mean value ± S.E.M. (*: p ≤ 0.05; **: p ≤ 0.01; ***: p ≤ 0.001).

Article Snippet: The membrane was incubated with antibodies diluted in Odyssey blocking buffer (LI-COR Biosciences) overnight at 4 °C to detect the following proteins: p-p38 MAPK (Tyr182; E-1; 1:500; Santa Cruz Biotechnology, Dallas, TX, USA), p38 MAPK (1:1000), p-Hsp27 (Ser82; 1:1000), Hsp27 (G31; 1:1000), AR (D6F11; 1:1000), HIF-1ɑ (1:1000; BD Biosciences, San Jose, CA, USA), eIF4E (1:1000; BD Biosciences), and beta-tubulin (1:10,000; Abcam, Cambridge, UK).

Techniques: Positive Control, Activation Assay, Western Blot, Control

Journal: Cancers

Article Title: p38 MAPK Inhibition Mitigates Hypoxia-Induced AR Signaling in Castration-Resistant Prostate Cancer

doi: 10.3390/cancers13040831

Figure Lengend Snippet: p38 MAPK inhibition prolongs survival of mice bearing CRPC xenografts. Subcutaneous V16D xenografts were established and treatment with vehicle or SB203580 (10 mg/kg) started when tumors reached 200 mm 3 , in a 5 days on and 2 days off schedule until endpoint. Kaplan-Meier survival curves are shown for the time to reach tumor volume endpoint (1000 mm 3 ) from treatment start. Tumor growth curves from individual mice are shown in .

Article Snippet: The membrane was incubated with antibodies diluted in Odyssey blocking buffer (LI-COR Biosciences) overnight at 4 °C to detect the following proteins: p-p38 MAPK (Tyr182; E-1; 1:500; Santa Cruz Biotechnology, Dallas, TX, USA), p38 MAPK (1:1000), p-Hsp27 (Ser82; 1:1000), Hsp27 (G31; 1:1000), AR (D6F11; 1:1000), HIF-1ɑ (1:1000; BD Biosciences, San Jose, CA, USA), eIF4E (1:1000; BD Biosciences), and beta-tubulin (1:10,000; Abcam, Cambridge, UK).

Techniques: Inhibition