Journal:

Article Title: Development of a Primary Tamarin Hepatocyte Culture System for GB Virus-B: a Surrogate Model for Hepatitis C Virus

doi:

Figure Lengend Snippet: Immunofluorescence staining for NS3 protein. Hepatocytes grown on glass coverslips were inoculated with GBV-B 3 days postplating and harvested 3 days postinfection unless noted otherwise. GBV-B NS3 protein was detected by immunofluorescence using a rabbit anti-GST-NS3 antiserum and goat anti-rabbit IgG-fluorescein. (A) Infected hepatocytes stained with anti-NS3 (magnification, ×100); (B) uninfected hepatocytes stained with anti-NS3 (×100); (C) infected hepatocytes stained with normal rabbit (prebleed; ×100); (D) lower magnification of infected hepatocytes stained with anti-NS3 (×50); (E) hepatocytes infected with a low multiplicity and harvested 21 days p.i. (×100); (F) higher magnification of infected hepatocytes stained with anti-NS3 (×200).

Article Snippet: Purified GST-NS3 protein (10 ng per well) was bound to Immulon 2 96-well plates (Dynatech Laboratories, Chantilly, Va.) in borate-buffered saline (145 mM NaCl, 6 mM NaOH, 48 mM H 3 BO 3 , 50 mM KCl [pH 8.2]).

Techniques: Immunofluorescence, Staining, Infection

Journal:

Article Title: Development of a Primary Tamarin Hepatocyte Culture System for GB Virus-B: a Surrogate Model for Hepatitis C Virus

doi:

Figure Lengend Snippet: Specificity of rabbit anti-GST-NS3 antiserum in immunofluorescence staining for NS3 protein. Hepatocytes were inoculated with GBV-B and harvested 3 days p.i. Cells were stained for NS3 protein using rabbit anti-GST-NS3 antiserum (A). To demonstrate specificity, the antiserum was adsorbed for 16 h at 4°C with 5 μg of purified GST (B) or NS3 (C) protein prior to being used for immunofluorescence staining. Adsorption with GST had no effect on staining, while adsorption with NS3 eliminated staining.

Article Snippet: Purified GST-NS3 protein (10 ng per well) was bound to Immulon 2 96-well plates (Dynatech Laboratories, Chantilly, Va.) in borate-buffered saline (145 mM NaCl, 6 mM NaOH, 48 mM H 3 BO 3 , 50 mM KCl [pH 8.2]).

Techniques: Immunofluorescence, Staining, Purification, Adsorption

Journal: Stem Cell Research & Therapy

Article Title: Anti-inflammatory treatment induced regenerative oligodendrogenesis in parkinsonian mice

doi: 10.1186/scrt124

Figure Lengend Snippet: Detrimental effect of chronic TNF-α treatment on mouse neural stem cell cultures . ( A ) Representative images showing NSC immunostaining for Nestin and Ki67 (counterstained with Hoechst); quantified in ( C ). ( B ) Images show representative immunostaining for TuJ1 in differentiated NSC cultures; quantified in ( D ). * P ≤0.05 Mann-Whitney-U-Test, error bars s.e.m.

Article Snippet: Osmotic mini-pumps (Alzet Osmotic Pumps, Cupertino, CA, USA, model 1002, 0.25 µl flow rate, brain infusion kit 3) for 14-day long TNF-α infusion were filled according to the manufacturer's protocol with either sterile saline (0.9% NaCl w/v) or with saline + 10 ng/ml TNF-α (Peprotech) under a sterile hood.

Techniques: Immunostaining, MANN-WHITNEY

Journal: Stem Cell Research & Therapy

Article Title: Anti-inflammatory treatment induced regenerative oligodendrogenesis in parkinsonian mice

doi: 10.1186/scrt124

Figure Lengend Snippet: Chronic ventricular TNF-α infusion negatively affects SVZ neurogenesis . (A ) Schematic showing the experimental setup. ( B ) Representative confocal maximum intensity projections showing TNF-α effect on EdU + and Dcx + cell numbers in SVZ (control: n = 6; TNF-α: n = 11) and pRMS (control: n = 7; TNF-α: n = 11); dotted line indicates ventricle surface; quantified in ( C ) and ( D ). # P ≤0.05 Student's t -test, * P ≤0.05 Mann-Whitney-U-Test, error bars s.e.m.

Article Snippet: Osmotic mini-pumps (Alzet Osmotic Pumps, Cupertino, CA, USA, model 1002, 0.25 µl flow rate, brain infusion kit 3) for 14-day long TNF-α infusion were filled according to the manufacturer's protocol with either sterile saline (0.9% NaCl w/v) or with saline + 10 ng/ml TNF-α (Peprotech) under a sterile hood.

Techniques: MANN-WHITNEY

Journal: PLoS ONE

Article Title: PPARγ Regulates Expression of Carbohydrate Sulfotransferase 11 ( CHST11/C4ST1 ), a Regulator of LPL Cell Surface Binding

doi: 10.1371/journal.pone.0064284

Figure Lengend Snippet: (A) Chip-Seq data of PPARγ and RXRα occupancy on the Chst11 gene according to Nielssen et al. . UCSC Genome Browser tracks at day 0, 1, 2 4 and 6 of differentiation are shown. Please note differences in y-axes. Two intronic PPARy-RXRα binding sites were designated site 1 and 2. (B) ChIP-PCR on 3T3-L1 preadipocytes and adipocytes. Chromatin was prepared on day 0 and day 6 of differentiation and subjected to immunoprecipitation using antibodies against PPARγ. Enriched DNA was analysed using quantitative PCR with primers located at site 1 and 2 in the Chst11 gene (dark gray and blacks bars, respectively). As a positive control, primers located at the Fabp4 PPREs (∼5500 bp from transcription start site; light gray bars) were used, primers located in the globin locus were used as a negative control (white bars). Results are shown relative to normalized ChIP recovery data of day 0 and results are representative of at least 3 independent experiments. (C and D) U2OS cells were cotransfected with a reporter construct (TKpGL3) containing Chst11 site 1 or site 2 sequences, or the parental reporter construct, together with empty (pCDNA) or PPARγ encoding expression vectors. Activation of the luciferase reporter in the absence or presence of 1 µM rosiglitazone is expressed as fold induction over that with empty reporter cotransfected with pCDNA in the absence of rosiglitazone after normalisation for Renilla luciferase activity. The error bars display SEM and significance is shown by the asterisks (p<0.05), n = 3 (E) Chst11 mRNA expression in 3T3-L1 adipocytes that had been treated with control or PPARy siRNA from the start of differentiation and analyzed at day 5. Relative mRNA expression levels were related to control siRNA treated cells and normalized for the TFIIb reference gene. The error bars display SEM and significance is shown by the asterisks (p<0.05), n = 3 (F) Chst11 mRNA expression in 3T3-L1 adipocytes and the effect of rosiglitazone treatment (1 µM, 24 h). Relative mRNA expression levels were normalized for the TFIIb reference gene. (G) As in panel F, but after treatment with the PPARγ antagonist GW9662 (10 µM, 24 h).

Article Snippet: In short, cells were transfected in 24-well plates with 1 µg reporter plasmid, 10 ng PPARy expression construct, and 2 ng pCMV-Renilla reporter plasmid (Promega).

Techniques: ChIP-sequencing, Binding Assay, Immunoprecipitation, Real-time Polymerase Chain Reaction, Positive Control, Negative Control, Construct, Expressing, Activation Assay, Luciferase, Activity Assay