Additional file 1: Figure S1. of Anti-cancer effect of pristimerin by inhibition of HIF-1α involves the SPHK-1 pathway in hypoxic prostate cancer cells

Pristimerin does not affect PI3K in PC-3 cells under hypoxia. PC-3 cells were treated with pristimerin (1 μM) and or SPHK-1 inhibitor (SKI) (10 μM) for 4 h under hypoxia. Effect of pristimerin on the expression of PI3K in hypoxic PC-3 cells. Western blotting was performed to determine the expression of PI3K and β-actin in hypoxic PC-3 cells. (TIF 66 kb

Epitope mapping and affinity measurement of two monoclonal antibodies (5H6, 3A5) against G core fragment.

<p>(A) Primary structure of G surface glycoprotein of RSV A2 strain and Gcf sequence. (B) Epitope mapping and affinity measurement of mAb (5H6 and 3A5) was performed using ELISA with known B-cell epitopes of the RSV G protein (peptide G/144-159, G/164-176, G/174-187, and G/190-204). The 96-well plates were coated with 200 ng or 100 ng of peptide corresponding to each epitope of the antibody. The wells were incubated with the indicated dilutions of mAbs.</p

Binding characteristics of monoclonal antibodies to Gcf derivatives with cysteine substitutions.

<p>(A) 96-well plates were coated with 50 ng of Gcf and mutant Gcf derivatives per well. Antibodies were serially diluted from 500 μg/ml and ELISA was performed. (B) Western blotting analysis with Gcf and mutant Gcf derivatives. For this, 100 ng and 3 μg of each indicated protein were loaded for blotting with 5H6 and 3A5, respectively.</p

Inhibition of Gcf-associated chemotactic activity by 5H6 and 3A5.

<p>Inhibition of <i>in vitro</i> chemotaxis activity of Gcf by mAbs was analyzed using chemotaxis assay with THP-1 cells. 10 μg of wild-type Gcf were pre-incubated with 100 μg of mAbs in the lower chamber, and 5 × 10⁵ THP-1 cells were added to the upper chamber later. Serum-free media alone or media with 10% FBS was used as a negative or positive control, respectively. The assembled plates were incubated at 37°C for 5 hr. Cells attached to bottom of the membrane were counted, and the percent migration was calculated.</p

Binding characteristics of monoclonal antibody to G proteins expressed by mammalian cells.

<p>(A) 96-well plates were coated with 200 PFU of purified RSV A2 virus particles. 5H6 was serially diluted from 10 μg/ml, and 3A5 from 500 μg/ml. The analysis was performed by using ELISA. (B) <i>In vitro</i> binding assay using flow cytometry. Binding of 5H6 or 3A5 to RSV particles was indicated by histograms and the mean fluorescence intensity (MFI). (C) Secreted G protein and Gcf were prepared from HEp-2 cells infected with RSV A2 and rAd/3XG, respectively. 96-well plates were coated with 5 μg of supernatant per well.</p

5H6 prevents weight loss primed by vvG.

<p>Effect of prophylactic administration of mAb 5H6 and 3A5 on weight change in BALB/c mice. BALB/c mice were scarified with 6×10⁶ PFU of recombinant vaccinia virus expressing RSV G protein (vvG). After 4 weeks, mice were intramuscularly administered with 300 μg of each mAb. All groups were challenged intranasally with 3 × 10⁶ PFU of RSV A2 1 day later (n = 4 mice/group) and then weighed each day. *, p < 0.05; **, p < 0.01; ***, p < 0.001, Significant increase on body weight for 5H6 or 3A5 treated group compared to PBS group.</p

<i>In vitro</i> Neutralization and <i>In vivo</i> RSV clearance activity of 5H6 and 3A5.

<p>(A) <i>In vitro</i> plaque reduction assay. Palivizumab, heparin, 5H6, and 3A5 were incubated with 200–300 PFU of RSV A2 for 1 hr at 37°C. Then, the mixture was added to HEp-2 cells for 1.5 hr, and the plaques were counted 5 days later. (B) Prophylactic treatment with mAb in mouse model. BALB/c mice were intramuscularly administered 100 μg of each mAb and challenged intranasally with 10⁶ PFU of RSV A2 1 day later (n = 4 mice/group). On day 4 post-infection, lung homogenates were prepared, and lung viral titers were measured by plaque assay. The limit of detection was 94 PFU/gram of lung tissue. N.D., not detected.</p

Additional file 2: Table S1. of Ethanol extract of Pinus koraiensis leaves containing lambertianic acid exerts anti-obesity and hypolipidemic effects by activating adenosine monophosphate-activated protein kinase (AMPK)

Composition of fat in HFD. (PDF 7 kb

Additional file 1: Figure S1. of Ethanol extract of Pinus koraiensis leaves containing lambertianic acid exerts anti-obesity and hypolipidemic effects by activating adenosine monophosphate-activated protein kinase (AMPK)

Flow diagram for the extraction and fractionation of LA from P. koraiensis. (TIF 2837 kb

Additional file 3: Figure S2. of Ethanol extract of Pinus koraiensis leaves containing lambertianic acid exerts anti-obesity and hypolipidemic effects by activating adenosine monophosphate-activated protein kinase (AMPK)

Food intake. (TIF 2565 kb