• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.91-94
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• 2000

The suspension cells of Taxus cuspidata were transformed with Agrobacterium tumefaciens harboring binary vector pCAMBIE1302 encoding mgfp. Transient transfection efficiency was compared by using the fluoremetric measurement. The transient transfection efficiency was improved by transformation with DMSO and/or sonication treatment. Optimum conditions for DMSO and sonication treatment were 3% and 30sec, respectively. selection and maintenance of transformed cells were continued for 3 months. An insertion of the mgfp gene in transformed cells was detected by PCR and an expression of GFP confirmed by the western blot analysis.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.427.2-428
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• 2002

Previously we formulated new cationic liposomes, DDC, composed of DOTAP. DOPE, and cholesterol (Chol) in 1 : 0.7 : 0.3 molar ratios, and showed that DDC efficiently deliver the plasmid DNA into ovarian cancer cell lines. Here, wild type p53 DNA was transfected into ovarian cancer cells, using the DOC as a nonviral vector and the expression and activity of p53 gene were evaluated in vitro and in vivo. The complexes of plasmid DNA (pp53-EGFP) and DDC were transfected into OVCAR-3 cells. The gene expression was determined by RT -PCR and western blot analysis. (omitted)

• Natural Product Sciences
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• v.19 no.3
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• pp.206-214
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• 2013

The crocetin derivatives, crocin (1), gentiobiosyl glucosyl crocetin (3), and mono-gentiobiosyl crocetin (4) were isolated from the fruit of Gardenia jasminoides (Gj) and crocetin (2) from the processed fruit of Gj (PGj) by column chromatography. Their structures were determined on the basis of spectroscopic methods including IR, MS, and NMR (1D and 2D). These compounds were evaluated for their inhibition activity on melanin production in ${\alpha}$-MSH (melanocyte stimulating hormone) activated B16F10 cells. Compounds 1 - 4 reduced melanin content in a dose-dependent manner at concentrations of 20 - 60 uM. They also suppressed tyrosinase protein and m-RNA expressions dose dependently, assayed by western blot analysis, and RT-PCR experiment in B16F10 murine melanoma cells.

• Natural Product Sciences
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• v.19 no.3
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• pp.258-262
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• 2013

Three ergosterol derivatives, ergosta-4,6,8(14),22-tetraen-3-one (1), ergosta-7,24(28)-dien-3-ol (2), and 5,8-epidioxyergosta-6,22-dien-3-ol(3) were isolated from the fruit body of Phellinus pini. Their structures were based on spectroscopic methods including IR, MS, and NMR (1D and 2D). These compounds were evaluated for their activity to decrease melanin production in ${\alpha}$-MSH (melanocyte stimulating hormone) activated B16F10 cells. Compound 1, 2, and 3 reduced melanin content in a dose-dependent manner at concentrations of 5~15 uM. They also suppressed the tyrosinase expression of protein and m-RNA level dose dependently by western blot analysis and RT-PCR experiment in B16F10 murine melanoma cells.

• BMB Reports
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• v.41 no.12
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• pp.875-880
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• 2008

Mbu-3 is a novel mouse brain unigene that was identified by digital differential display. In this study, expression of the gene was chased through developmental stages and the protein product was identified in the brain. The cDNA sequence was 3,995-bp long and contained an ORF of 745 AA. Database searches revealed that the chicken SST273 gene containing LRR- and Ig-domain was an mbu-3 orthologue. Tissue specificity for the gene was examined in embryos and in brains at post-natal and adult stages. During the embryonic stages, mbu-3 was localized to the central nervous system in the brain and spinal cord. In the early post-natal stages, the gene was evenly expressed in the brain. However, with aging, expression was confined to specific regions, particularly the hippocampus. The protein was approximately 95 kDa as determined by Western blot analysis of brain extracts.

• Journal of Veterinary Science
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• v.25 no.4
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• pp.54.1-54.16
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• 2024

Importance: As one of the main etiologic agents of infectious diseases in pigs, pseudorabies virus (PRV) infections have caused enormous economic losses worldwide. EP0, one of the PRV early proteins (EP) plays a vital role in PRV infections, but the mechanisms are unclear. Objective: This study examined the function of EP0 to provide a direction for its in-depth analysis. Methods: In this study, the EP0-deleted PRV mutant was obtained, and Tandem Mass Tag-based proteomic analysis was used to screen the differentially expressed proteins (DEPs) quantitatively in EP0-deleted PRV- or wild-type PRV-infected porcine kidney 15 cells. Results: This study identified 7,391 DEPs, including 120 and 21 up-regulated and down-regulated DEPs, respectively. Western blot analysis confirmed the changes in the expression of the selected proteins, such as speckled protein 100. Comprehensive analysis revealed 141 DEPs involved in various biological processes and molecular functions, such as transcription regulator activity, biological regulation, and localization. Conclusions and Relevance: These results holistically outlined the functions of EP0 during a PRV infection and might provide a direction for more detailed function studies of EP0 and the stimulation of lytic PRV infections.

• Biomedical Science Letters
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• v.15 no.2
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• pp.141-146
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• 2009

Paclitaxel, an antimicrotubule agent, binds to beta-tubulin in the microtubule and stabilizes the polymer, thereby repressing dynamic instability. Here, we have demonstrated that microtubule cytoskeletal architecture involved in regulation of the COX-2 expression in chondrocyte treated with paclitaxel. Paclitaxel enhanced COX-2 expression and prostaglandin E2 production, as indicated by the Western blot analysis, reverse transcriptase PCR(RT-PCR) and immunofluorescence staining, and $PGE_2$ assay, respectively. In our previous data have shown that paclitaxel treatment stimulated activation of ERK-1/2 and p38 kinase(Im et al., 2009). SB203580, an inhibitor of p38 kinase, blocked the induction of COX-2 expression by paclitaxel. Also PD98059, an inhibitor of ERK-1/2 kinase was blocked the induced COX-2 expression. These results indicate that activation of ERK-1/2 and p38 kinase is required for COX-2 expression induced by paclitaxel in rabbit articular chondrocytes.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.533-539
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• 2011

Sulfuretin is one of the main flavonoids produced by Rhusverniciflua. Sulfuretin has been shown to exhibit many pharmacological activities including anti-oxidant, anti-obesity, anti-inflammatory and anti-mutagenic activities. However, the anti-skin photoaging effects of sulfuretin has not yet been reported. In the present study, we investigated the inhibitory effect of sulfuretin on the expression levels of MMP-1 and -3 in the human dermal fibroblast cells. Western blot analysis and real-time PCR revealed sulfuretin inhibited UVB-induced MMP-1 and -3 expressions in a dose-dependent manner. UVB-induced MAPK/NF-${\kappa}B$/p50 activation and MMP expression were completely blocked by pretreatment of sulfuretin. Taken together, sulfuretin could prevent UVB-induced MMP expressions through inhibition of MAPK/NF-${\kappa}B$/p50 activation.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.4
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• pp.455-460
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• 1998

The present study was aimed at investigating whether the development of hypertension is related with an altered expression of nitric oxide synthases (NOS) in the kidney. By Western blot analysis, the expression of bNOS and ecNOS isoforms was determined in the kidney of deoxycorticosterone acetate (DOCA)-salt and two-kidney, one clip (2K1C) rats. In DOCA-salt hypertension, the expression of both bNOS and ecNOS was decreased, along with tissue contents of nitrites. In 2K1C hypertension, the nitrite content of the clipped kidney was decreased along with ecNOS levels, whereas neither the nitrite content nor the expression of NOS isoforms was significantly altered in the contralateral non-clipped kidney. These results suggest that the development of hypertension is associated with an altered renal expression of NOS and nitric oxide generation in DOCA-salt and 2K1C rats.

• Journal of Pharmacopuncture
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• v.9 no.1
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• pp.75-82
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• 2006

Objective : This research was carried out for the development of medicine for vitiligo treatment and focused on the effect of Hominis Placenta extract on Melanin synthesis of B16 melanoma cells. Methods : Acitivity of tyrosinase playing a vital role in synthesis of Melanin and the quantity of Melanin, which is the final product in cultured B16 melanoma cells, effects of Hominis Placenta extract were measured. Results: The results indicated that Hominis Placenta extract increased both the amount of Melanin and the activity of tyrosinase according to the concentration, and they also supported by western blot analysis. Conclusion : The results suggests that Hominis Placenta extract has an advantageous effect on the promotion of Melanin synthesis and will contribute to the development of vitiligo treatment through further related studies.