Journal: British journal of haematology

Article Title: Hypomethylating agent alters the immune microenvironment in AML and enhances the immunogenicity of a DC/AML vaccine

doi: 10.1111/bjh.15818

Figure Lengend Snippet: MOLM-14 (A) and MV4–11 (B) cells were exposed to decitabine (DAC) at 25nM or 50nM daily for 3 days and analyzed for ERV-3 expression using western blotting on day 4 of treatment (WB). The results showed significant upregulation of ERV-3 expression following treatment. Due to activation of the ERV pathway, increased IFNß expression was observed with WB analysis.

Article Snippet: Cells were assessed for ERV-3 (anti-ERV-3 Everest Biotech, Oxfordshire, UK, 30 kD) and IFNβ (anti-IFNβ Abcam, Cambridge, UK, 22 kD) expression.

Techniques: Expressing, Western Blot, Activation Assay

Journal: Proteome Science

Article Title: Identification of differentially regulated proteins in a patient with Leber's Congenital Amaurosis – a proteomic study

doi: 10.1186/1477-5956-5-5

Figure Lengend Snippet: Identification of differentially regulated proteins in LCA retina

Article Snippet: After transfer by electroelution to nitrocellulose Hybond C-extra membranes, blots were blocked with 5% milk, 5% fetal calf serum, and 0.05% Tween-20 in 80 mM Na 2 HPO 4 , 20 mM NaH 2 PO 4 , 100 mM NaCl, pH 7.5 (PBS-T), and incubated overnight at 4°C with anti-triosephosphate isomerase (Everest Biotech, Oxfordshire, UK) diluted 1:1,000, anti-αA-crystallin (Acris Antibodies, Hiddenhausen, Germany) diluted 1:1,000, anti-β-tubulin (Abcam, Cambridge, UK) diluted 1:1,000, and anti-ATP synthase subunit β (Abcam, Cambridge, UK) diluted 1:500, anti-β-actin (Abcam, Cambridge, UK) diluted 1:100.

Techniques:

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Command and Compensation in a Neuromodulatory Decision Network

doi: 10.1523/JNEUROSCI.3707-11.2012

Figure Lengend Snippet: A, CNS wholemounts from newly eclosed flies expressing either TRPM8 alone (left), or TRPM8 plus an RNAi construct directed against the bursicon subunit pburs (right) in CCAP-expressing neurons, were labeled with an anti-pburs antibody. pburs-RNAi expression substantially reduces anti-pburs immunostaining in both groups of bursicon-expressing neurons, the BSEG (arrowheads) and the BAG (brackets). Scale bar, 50 μm. B, Bar graphs showing that flies expressing pburs-RNAi did not expand their wings (top). Unlike control flies not expressing pburs-RNAi, approximately 30% of flies in which pburs levels are knocked down exhibit expansional behavior (middle), but only after environmental delays in the time till abdominal flexion (bottom), even when the CCAP-expressing neurons are stimulated (ΔTemp). C, Burs-Gal4 drives UAS-TRPM8 expression in both subsets of bursicon-expressing neurons, the BAG and the BSEG, as revealed by double-labeling with anti-burs (magenta) and anti-TRPM8 (green) immunolabeling. Scale bar, 50 μm. D, Box plots show that under adverse conditions, a 15 min temperature shift to 18°C from 25°C (ΔTemp) rapidly induces wing expansion in Burs-Gal4>TRPM8 flies (+) compared with control animals either of the same genotype and not subjected to temperature shift (-) or lacking the Burs-Gal4 driver (Con). The no temperature shift control plot includes two animals that tonically flexed their abdomens, but did not fully expand their wings. As noted in Materials and Methods, this sometimes happens after prolonged delays. In these cases, “Expansional Delay” indicates the time until termination of abdominal flexion. *** and NS, indicate p<0.001 and p>0.05 as evaluated by t-test. Number of animals in each group is in parentheses above each plot.

Article Snippet: Immunohistochemistry and immunoblotting Whole mounts of the central nervous system from either males or females were prepared, stained, and analyzed by confocal microscopy as described previously ( Luan et al., 2006b ) using the following primary antibodies: rabbit anti-burs (AKA, anti-bursicon α-subunit; 1: 5000), rat anti-pburs (AKA, anti-bursicon β-subunit; kind gift of Aaron Hsueh, used at 1: 500 ), goat anti-TRPM8 (Everest Biotech, Oxfordshire, UK, used at 1:2000).

Techniques: Expressing, Construct, Labeling, Immunostaining, Immunolabeling

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Command and Compensation in a Neuromodulatory Decision Network

doi: 10.1523/JNEUROSCI.3707-11.2012

Figure Lengend Snippet: A, Box plots showing the time required to complete expansional behaviors (i.e. Expansional Delay) for flies expressing TRPM8 in the indicated cell group (Driver) with or without stimulation (ΔTemp). (-)DBD, controls lacking the BursGalDBD-U6A1 hemidriver. As noted in the legend of Fig. 1, some flies that attempted expansion after prolonged delays failed to expand their wings, in which case the expansional delay represents the time they took to complete abdominal flexion. The number of flies tested in each group is indicated above the graph, as are the results of t-test comparisons between groups. NS, p>0.05; ***, p<0.001. B, Western blots showing blood titers of bursicon (i.e. anti-burs immunostaining) resulting from stimulation (ΔTemp) of the BAG (top) or BSEG (bottom). PC, burs positive control indicates release during normal expansion. Volumes of blood loaded per sample are indicated. C, Wing morphology (arrows) and tanning (arrowheads) of flies expressing TRPM8 in either the BAG or BSEG evaluated 3 h after eclosion. D, Abdominal nerves from BSEG>TRPM8 animals with (right) or without (left) BSEG stimulation immunostained for bursicon using anti-burs antibodies. Scale bars, 50 μm. E, Box plot showing quantified burs-IR data from abdominal nerves of the indicated number of animals of each kind shown in D, demonstrating release of bursicon from these nerves in response to BSEG stimulation. mpi; mean pixel intensity (background-subtracted). **, p<0.01 by t-test.

Article Snippet: Immunohistochemistry and immunoblotting Whole mounts of the central nervous system from either males or females were prepared, stained, and analyzed by confocal microscopy as described previously ( Luan et al., 2006b ) using the following primary antibodies: rabbit anti-burs (AKA, anti-bursicon α-subunit; 1: 5000), rat anti-pburs (AKA, anti-bursicon β-subunit; kind gift of Aaron Hsueh, used at 1: 500 ), goat anti-TRPM8 (Everest Biotech, Oxfordshire, UK, used at 1:2000).

Techniques: Expressing, Western Blot, Immunostaining, Positive Control

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Command and Compensation in a Neuromodulatory Decision Network

doi: 10.1523/JNEUROSCI.3707-11.2012

Figure Lengend Snippet: Summary of Manipulations and Results

Article Snippet: Immunohistochemistry and immunoblotting Whole mounts of the central nervous system from either males or females were prepared, stained, and analyzed by confocal microscopy as described previously ( Luan et al., 2006b ) using the following primary antibodies: rabbit anti-burs (AKA, anti-bursicon α-subunit; 1: 5000), rat anti-pburs (AKA, anti-bursicon β-subunit; kind gift of Aaron Hsueh, used at 1: 500 ), goat anti-TRPM8 (Everest Biotech, Oxfordshire, UK, used at 1:2000).

Techniques: Activation Assay