Journal: Scientific Reports

Article Title: Fibroblast growth factor (FGF) signaling regulates transforming growth factor beta (TGF β )-dependent smooth muscle cell phenotype modulation

doi: 10.1038/srep33407

Figure Lengend Snippet: HASMCs were cultured in the growth medium (M231 + SMGS). ( A ) ELISA quantification of TGF β 2 present in the extracellular culture medium of control and FRS2 α knockdown HASMCs. Data represent mean ± SD (* P < 0.05 compared to control; unpaired two-tailed Student’s t test. N = 3). ( B – F ) Upper panels: Immunoblot analysis of TGF β signaling in control, EGFR, FGFR1, FRS2 α , IGF1R, and PDGFR β knockdown HASMCs. HASMCs were serum starved for 8 hr then stimulated with TGF β 1 (0.5 ng/ml) for different time points. Blots are representative of three independent experiments. Bottom panels: Band intensities of p-Smad2 were normalized to Smad2 and expressed as a fraction of a control value.

Article Snippet: Thereafter, the respective extracellular medium was collected and a commercially available sandwich ELISA kit (R&D Systems #DB250) was used to evaluate the secretion of TGF β 2 in the extracellular medium.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Control, Knockdown, Two Tailed Test, Western Blot

Journal:

Article Title: MILD CHOLESTEROL DEPLETION REDUCES AMYLOID-? PRODUCTION BY IMPAIRING APP TRAFFICKING TO THE CELL SURFACE

doi: 10.1111/j.1471-4159.2009.06126.x

Figure Lengend Snippet: Cholesterol depletion reduces Aβ, APP CTFs and cell surface APP without affecting AICD generation. A) PS70 or HEK cells were treated with 20 μM lovastatin in the presence or absence of DLFBS for 48 hours and Aβ levels were measured by sandwich ELISA. Treatment with lovastatin led to a reduction of Aβ40 and Aβ42 that is prevented by the presence of cholesterol-containing serum. B) PS70 cells were treated with lovastatin or MβCDX in the presence or absence of DLFBS for 48 hours and membrane preparations were obtained as described (Sastre et al. 2001). Levels of full-length APP and APP CTFs were measured by Western blot. Cholesterol depletion reduced both α- and β-CTFs that can be recovered by addition of cholesterol-containing serum. Quantification from four independent experiments is shown below. C) HEK cells were transfected with BAP-APP construct and treated with vehicle, lovastatin, 5 or 10 mM MβCDX in FBS or DLFBS. Cell surface APP was biotinylated and the cell lysate was subjected to Western blot analysis for APP expression. Treatment with lovastatin or/and MβCDX reduced cell surface APP without altering total APP expression. D) Cell-free AICD generation assay. PS70 cells were treated with different concentrations of MβCDX and membrane preparations were incubated at 37 °C for 2 hours. Extent of cholesterol reduction is indicated below. APP CTFs and AICD were measured by Western blot. No changes in AICD generation were observed after cholesterol depletion. Incubation at 4 °C or with DAPT inhibited the production of AICD. Average data from three independent experiments are shown below. E) Aβ1-x levels were measured in the same cell-free assay and results were normalized to levels of APP CTFs. Only a slight decrease was observed at the highest MβCDX concentration. Incubation at 4°C and more markedly treatment with DAPT reduced Aβ generation. Data represent the average of two independent experiments.

Article Snippet: To measure human Aβ 1-42 and Aβ 1-x we used sensitive ELISA kits (Wako, Japan and IBL, Germany, respectively).

Techniques: Sandwich ELISA, Western Blot, Transfection, Construct, Expressing, Incubation, Cell-Free Assay, Concentration Assay

Journal: PLoS ONE

Article Title: Gender difference in NASH susceptibility: Roles of hepatocyte Ikkβ and Sult1e1

doi: 10.1371/journal.pone.0181052

Figure Lengend Snippet: Wild type (Wt), myeloid IKKβ deficient ( IkbkbΔmye ), or hepatocyte IKKβ deficient ( IkbkbΔhep ) mice in both genders, were fed regular chow (Control) or high cholesterol and saturated fat diet (HCFD) from 2.5 months of age for 20 weeks. (A) Immunoblotting of whole liver lysate and (B) densitometric analysis of pAMPK/AMPK, nSREBP-1c, PPARδ, pJNK-1/JNK-1, and pJNK-2/JNK-2. (C) Plasma adiponectin levels measured by ELISA. (D) Real-time PCR results of PPAR α, δ, and γ genes. *p<0.05, **p<0.01, ***p<0.001 compared to Control diet within gender and within genotype. †p<0.05, ††p<0.01 compared to other genotype within gender and within diet.

Article Snippet: Plasma ALT, insulin and adiponectin concentrations were measured by ELISA kits (ALT: Sigma Diagnostic, St. Louis, MO, insulin: Shibayagi Co., Gunma, Japan, adiponectin: ALPCO Diagnostics, Salem, NH, respectively).

Techniques: Control, Western Blot, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Clinical Microbiology

Article Title: Evaluation of In-House and Commercial Serological Tests for Diagnosis of Human Tularemia

doi: 10.1128/JCM.01440-17

Figure Lengend Snippet: ROC curves for detection of IgM or IgG anti-F. tularensis antibodies using immunofluorescence assays (IFAs) or ELISAs. AUC, area under the curve.

Article Snippet: The sensitivities and specificities of the tests evaluated were, respectively, 97% and 91.5% for the Serazym ELISAs, 96.3% and 96.8% for the Serion ELISA-IgG tests, 94.8% and 96.8% for the Serion ELISA-IgM tests, and 97% and 84% for the VIRapid tularemia tests.

Techniques: Immunofluorescence