Protein levels differentially expressed in the striatum.

<p>The levels were illustrated as the relative normalized volumes (RNV) of the proteins. Bars represent the means ± standard deviation (n = 3 at each group). *<i>p</i> < 0.05, ** <i>p</i> < 0.01 and *** <i>p</i> < 0.001. Arrowheads represent protein spots.</p

Protein profiles with two-dimensional electrophoresis.

<p>(A) Striatal tissue proteins obtained over different pI ranges. (B) Identified protein spots in gels. Differentially-expressed protein spots are circled with green. (C) Cluster matrix analysis. Red corresponds to high expression and green corresponds to low expression.</p

Results of the pole test.

<p>MPTP administration induced an increase in the descending time of mice, which was restored by oral administration of Korean Red Ginseng (KRG). Data are presented as the means ± the standard deviation. **<i>p</i> < 0.01. Saline, vehicle injected group; MPTP, MPTP injected group; MPTP+KRG, MPTP injected and KRG administrated group.</p

List of differentially expressed protein spots following MPTP and KRG administration.

<p>List of differentially expressed protein spots following MPTP and KRG administration.</p

KEGG pathway list of 12 proteins which were down-regulated in MPTP versus saline and up-regulated in MPTP+KRG versus MPTP.

<p>KEGG pathway list of 12 proteins which were down-regulated in MPTP versus saline and up-regulated in MPTP+KRG versus MPTP.</p

Expression of tyrosine hydroxylase in the nigrostriatal pathway on day 3.

<p>MPTP injection induced dopaminergic neuronal death in the substantia nigra (A) and the striatum (B), but KRG administration suppressed it. Scale bar, 200 μm. Data are presented as the means ± standard deviation. *<i>p</i> < 0.05 and **<i>p</i> < 0.01.</p

Caffeic Acid Phenethyl Ester Inhibits Alpha-Melanocyte Stimulating Hormone-Induced Melanin Synthesis through Suppressing Transactivation Activity of Microphthalmia-Associated Transcription Factor

Caffeic acid phenethyl ester (<b>1</b>), a natural compound found in various plants and propolis, is a well-known anti-inflammatory, immunomodulatory, and cytotoxic agent. The present study aimed to investigate the molecular events underlying the antimelanogenic activity of <b>1</b> in alpha-melanocyte stimulating hormone (α-MSH)-stimulated B16-F10 melanoma cells. In this investigation, <b>1</b> effectively reduced α-MSH-stimulated melanin synthesis by suppressing expression of melanogenic enzymes such as tyrosinase, tyrosinase-related protein-1 (TRP-1), and tyrosinase-related protein-2 (TRP-2), although this compound did not directly inhibit tyrosinase enzyme activity. On the other hand, the expression and nuclear translocation of microphthalmia-associated transcription factor (MITF) as a key transcription factor for tyrosinase expression regulating melanogenesis were not affected by treatment with <b>1</b>. The upstream signaling pathways including cAMP response element-binding protein (CREB), glycogen synthase kinase-3β (GSK-3β), and Akt for activation and expression of MITF were also not influenced by <b>1</b>. Interestingly, <b>1</b> inhibited transcriptional activity of a tyrosinase promoter by suppressing the interaction of MITF protein with an M-box containing a CATGTG motif on the tyrosinase promoter. Given the important role of MITF in melanogenesis, suppression of <b>1</b> on the function of MITF to transactivate tyrosinase promoter may present a novel therapeutic approach to treat hyperpigmentation disorders

AF inhibits iNOS and COX-2 mRNA and protein levels in RAW 264.7 macrophage cells.

<p>RAW264.7 macrophage cells were treated with 0–50 mM AF and then co-treated with LPS for 24 h. Protein (A) and mRNA (B) levels were determined by RT-PCR and Western blot analysis, respectively. The data shown are representative of three independent experiments and indicate the mean ± SEM. *p < 0.05 and **p < 0.01 indicate significant differences from the LPS alone-treated cells.</p

Effects of AF on LPS-induced nuclear translocation of NF-κB and AP-1 in RAW 264.7 macrophage cells.

<p>Cells were treated in the presence or absence of various concentrations (0–50 μg/ml) of AF for 30 min and further incubated with or without 100 ng/ml of LPS for 15 min. Then, the cells were harvested and separated into nuclear (A) and cytosolic extracts (B), as described in the Methods section. The protein extracts were separated on SDS-PAGE gels and immunoblotted using Western blot analysis. The antibodies used were anti-NF-κB, -p-c-Jun, -hnRNP, -IκB, and -β-actin. hnRNP and β-actin were used as controls for nuclear- and cytosol-specific proteins, respectively. (C), (D) EMSA showing the reduction in NF-κB and AP-1 DNA binding activity in nuclear proteins. Cells were prepared by stimulation with 100 ng/mL LPS for 15 min and pre-treated with 50 μM AF for 30 min. (E), (F) The translocation of NF-κB (p65) and AP-1 to the nucleus was analyzed by confocal microscopy. Macrophages were immunostained using FITC for NF-κB and AP-1 and Hoechst to label nuclei. White scale bars, 5 μm. NT, no treatment; A, AF; L, LPS; L+A, LPS with AF.</p

Suppression of apoptotic signal events by GM3 RNA interference in CDDP-exposed cells.

<p>HCT116 cells were transfected with three siRNA targeting GM3 and negative control siRNA. Cells were then cultured for 12(30 µg/mL). Total RNA and protein lysates from the cells were prepared as described in Materials and Methods. (A) The levels of GM3 synthase mRNA in total RNA obtained from each cell was detected by RT-PCR. (B) Total protein was prepared from the cells and analyzed by western blotting using the indicated antibodies.</p