• Title/Summary/Keyword: 단백질제거작용

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Studies on Whitening and UV Damage Protection Effect of the Pterin Compounds (테린계 화합물의 미백 및 자외선 손상방어 효능에 관한 연구)

  • Kim, Nam-Kyoung;Kim, Mi-Hwa;Kang, Nae-Im;Choi, Shin-Wook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.1 s.60
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    • pp.53-60
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    • 2007
  • (6R)-5,6,7,8-tetrahydrobiopterin ($6-BH_4$) cofactor is essential for various process, and is present in probably every cell or tissue of higher organism. $6-BH_4$ is required lot various enzyme activities, and for less defined functions at the cellular level. And it is well known about the antioxidant effects as a non-protein compound. Recently, scientists proposed another roles for $6-BH_4$ in melanogenesis. $6-BH_4$ is a well known tyrosinase inhibitor. In this study, we found that methyl-$BH_4$ and $6-BH_4$ have antioxidant activities and inhibitory activity for melanin synthesis. These pterin compounds were not toxic in HaCaT and B16F10 cells and showed scavenging activity against DPPH radicals. We also showed that pterin compounds decreased protein levels of tyrosinase and TRP-1. In a clinical test, pterin compounds showed the significant skin whiteining effect after treatment for 3 weeks. Furthermore pterin compounds significantly suppressed the UVB-induced expression of $PGE_2$ and IL-6 genes induced UVB In HaCaT and inhibited UVB-induced melanogenesis in B16F10 cells. These results showed the effect of pterin compounds as a cosmeceutical ingredient.

The Overexpression of Subtilisin Enzyme Using Mutations on Transition State Regulatory Proteins of AprE Promoter and Development of Bacillus subtilis Host System (AprE Promoter전이상태 조절인자 변이주를 이용한 공업적 효소의 과발현과 고초균 숙주계의 개발)

  • 류성호;박승환김병기
    • KSBB Journal
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    • v.11 no.1
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    • pp.8-14
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    • 1996
  • Bacillus subtillis strains with transition state regulator mutations and a spore mutation were developed for the overexpression of apsE and for the enhancement of expression level. Among the many regulator genes, degU and hpr were chosen as a representative positive and negative regulator for the aprE, respectively. Spo II G was used for the construction of asporogeneous strains. All the mutants were constructed from two protease-deleted strain DB104 and the apsE gene was transformed with an integration vector pMK101. DB104(deg$U^h$(32) $his^+$)::pMK101(Cm) and DB104($\Delta$her(Em))::pMKl01(Cm) show 7-fold and about 2-fold increase in aprE expression level, respectively. But the effect of transition state regulator mutation on the aprE expression was diminished when the integrated aprE gene was amplified by the high concentration of chloramphenicol, i. e. 30 $\mu\textrm{g}$/ml. DB104($\Delta$spoIIG(Pm) degUh(32) his+)::pMK101(Cm) and DB104($\Delta$spoIIG(Pm) $\Delta$hpr(Em))::pMK101 double mutant show 10-fold and 3-fold increase in aprE expression level, respectively. The results suggest that sporulation mutation and transition state regulator mutation have independent and additive effect on the aprE expression, and the same gene dosage effect on the transition state regulator mutation was also identified.

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Inhibition of Plaque Formation on the Titanium Surface by Anti-bacterial Varnish (항균제 처리한 titanium 표면의 치태형성 억제효과)

  • Chung, Hyun-Ju;Lee, Sang-Hyun;Kim, Yung-Jun;Williams, Ray C.
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.707-727
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    • 2000
  • 매식된 인공치아의 성공을 위해서는 적절한 교합과 수동적 적합성을 갖는 보철물의 제작과 구강내 노출 직후부터의 세균성 치태조절이 요구된다. 본 연구는 전처리(passivation과 tridodecyl - methyl - ammonium chloride(TDMAC) 처리)가 다른 타이타늄 표 면에 chlorhexidine varnish와 테트라사이클린 을 도포시 약제의 방출역학을 알아보고 구강내 치태형성의 억제정도를 평가하기 위하여 시행 되었다. 이를 위해 방출용액으로 인산완충액 성분의 인조타액을 1일${\sim}$1개월간 매일 교환하여 약제농도를 측정하고 타이타늄 박막에 잔류한 약제 활성을 측정하였으며 항균제 도포한 타이타늄 원판을 부착한 장치를 구강내 위치시킨 1일${\sim}$3주 후 원판을 제거하여 주사전자현미경으로 세균 부착상을 관찰하였다. 테트라사이클린은 TDMAC 처리된 표면에서 $10{\sim}18$일까지 유효농도로 방출되었고 표면의 유효 항균 활성은 $3{\sim}4$주간 유지되었으며, chlorhexidine varnish 도포 시에는 TDMAC 전처리시 초기에 $3{\sim}7$일 간 증가한 유효 항균 활성을 방출하여 매식지대치 등에 이러한 항균제도포 시 매식치 주위환경에 항균활성 공급원으로 작용할 수 있음을 보였다. 주사현미경적 관찰시 모든 타이타늄 표면에서 구강내 위치 30분 후에는 세균이 부착되어 있지 않고 타액 단백질 성분에서 유래한 것으로 보이는 피막물질이 표면을 부분 또는 전면에 걸쳐 덮고 있었다. 구강내 노출 2시간 후 항균제 미도포 표본들에는 약간의 구균이 단층으로, $1{\sim}3$일 후에는 부분적으로 두꺼운 세균층을 형성하였고 7일 후에는 표면전체에 걸쳐 세균층이 덮여있었으며 주로 구균과 약간의 간균이 주종을 이루었다. 항균제 도포시 구강내 노출 1주일 이전까지는 미도포군에 비해 치태형성이 지연되는 경향을 보였지만 2주 이후에는 세균 수나 치태형성 양상이 유사하였다. 이 연구로부터 항균제 도포시 1주일 이전의 초기 치태형성을 감소시킬 수 있음을 알 수 있었으며 이러한 연구결과는 타이타늄 임프란트 지대치 표면에 항균제의 도포가 임상적으로 유용할 수 있음을 시사하였다.

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Rice Cell Origin Recombinant Human Granulocyte Macrophage Colony-Stimulating Factor (rrhGM-CSF) Could Improve the Wound Healing in Diabetic Hamster (당뇨가 유발된 햄스터 창상치유에 미치는 벼세포 유래 GM-CSF의 효과)

  • Han, Kyu-Boem;Heo, Si-Hyun;Jeong, Jin-Ju;Han, Man-Deuk;Kim, Wan-Jong;Shin, Kil-Sang
    • Applied Microscopy
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    • v.39 no.3
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    • pp.253-260
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    • 2009
  • GM-CSF is a multipotent growth factor, which also plays an important role during the process of wound healing. rrhGM-CSF was specifically produced from rice cell culture in our laboratory (Hanson Biotech Co., Ltd, Daejeon). The rrhGMCSF contains more oligosaccharide side chains than any other types of GM-CSF. This work was taken to evaluate the influence on wound healing of rrhGM-CSF in male golden hamsters. Full thickness skin defects of 9 mm in diameter were made in the back of hamsters, and 100 ${\mu}L$ ointment containing rrhGM-CSF 50 ${\mu}g/mL$ was applied. Control groups were given ointment without rrhGM-CSF. The wound sizes were relatively reduced and skin was well regenerated in the experimental group compared with the control group. Structurally, reepithelialization and architecture of the skin following injury were well accomplished in the experimental group. And also, positive reaction of PCNA of the skin following injury was more prominent in rrhGM-CSF containing ointment treatment group. Since this type of GM-CSF has highly glycosylated side chains, the effectiveness might be retain longer and stable, regarding acceleration of wound healing in the animal model. The present study has important implications for further development of the therapeutic manipulation of wound healing using rrhGM-CSF.

Antioxidative Effects of Mugwort (Artemisia vulgaris L.) Extracts Diet on ICR Mouse Skin (애엽 추출물 섭취가 ICR 마우스의 피부조직에 미치는 항산화 효과)

  • Park, Si-Hyang;Cho, Duck-Moon;Choi, Gyeong-Lim;Choi, Yeung-Joon;Choi, Jin-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.12
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    • pp.1523-1528
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    • 2007
  • The feeding effects of mugwort (Artemisia vulgaris L.) extracts (ME) on the anti-oxidative actions of ICR mouse skin was investigated. To study the antioxidative effects of ME on ICR mouse skin, female ICR mice were grouped into basic diet group (control), ascorbic acid diet group (AA-2.5, AA-5.0, AA-10.0 and AA-20.0 mg/kg BW/day) as a positive control and experimental diet group (mugwort extract; ME-25, ME-50, ME-100, and ME-200 mg/kg BW/day) and fed for 10 weeks. Protein contents in ME-50, ME-100, and ME-200 feeding group were increased ($3.1%{\sim}11.1%$) and hydroxyl radical contents were significantly decreased ($10.4%{\sim}17.4%$) compared to control group. Oxidative stress signals and oxidized protein contents were significantly reduced to the range of 15.3 to 17.1% in ME-100 and ME-200 groups. Also, superoxide dismutase (SOD) activity was significantly increased to the range of 15.0% to 23.3% in ME-100 and ME-200 groups. Catalase activities were significantly increased ($14.0%{\sim}36.9%$) in all groups in a dose-dependent pattern. Antioxidative ability of ME showed similarity to that of ascorbic acid.

Analysis of coenzyme Q10 in human plasma by high performance liquid chromatography (고성능액체크로마토그라피를 이용한 혈장 내 코엔자임 큐텐 분석)

  • Park, Yong-Sun;Park, Sang-Boem;Song, Sean-Mi;Kim, Yong-Woo;Lee, Kyoung-Ryul
    • Analytical Science and Technology
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    • v.22 no.6
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    • pp.514-518
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    • 2009
  • Coenzyme $Q_{10}$($CoQ_{10}$), a vitamin E-like substance, represents a components of the complex antioxidant system of the human organism. $CoQ_{10}$ levels in human plasma were determined by high performance liquid chromatography (HPLC) with UV detection. It was dissociated from lipoproteins by methanol and extracted into n-hexane with liquid-liquid extraction procedure, after centrifugation, the supernatant was dried under nitrogen gas stream. The residue was dissolved in the absolute ethanol. Determination of $CoQ_{10}$ was performed on a $C_{18}$ reversed-phase analytical column with ultraviolet detection at 275 nm and the mobile phase containing 15% (v/v) ethanol in methanol at a flow rate of 1.7 mL/min. The low limit of quantitation was 0.02 mg/L (S/N=10), the linearity between the concentration and peak height is from 0.1 to 2.0 mg/L. Twenty-four randomly selected plasma samples from apparently healthy, 27 to 44 year old individuals (males and females) were analyzed for total $CoQ_{10}$. The average level in these subjects was $0.62{\pm}0.13mg/L$ with the range of 0.41-0.98 mg/L. This method has a specific and a sufficient limit of quantitation (LOQ) for analysis of $CoQ_{10}$ in human plasma in both a clinical study and research at laboratories.

Microtubule-damaging Chemotherapeutic Agent-mediated Mitotic Arrest and Apoptosis Induction in Tumor Cells (미세소관-손상 항암제 처리에 의한 세포주기의 정지 및 에폽토시스 유도)

  • Jun, Do Youn;Kim, Young Ho
    • Journal of Life Science
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    • v.26 no.3
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    • pp.376-386
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    • 2016
  • Apoptosis induction has been proposed as an efficient mechanism by which malignant tumor cells can be removed following chemotherapy. The intrinsic mitochondria-dependent apoptotic pathway is frequently implicated in chemotherapy-induced tumor cell apoptosis. Since DNA-damaging agent (DDA)-induced apoptosis is mainly regulated by the tumor suppressor protein p53, and since more than half of clinical cancers possess inactive p53 mutants, microtubule-damaging agents (MDAs), of which apoptotic effect is mainly exerted via p53-independent routes, can be promising choice for cancer chemotherapy. Recently, we found that the apoptotic signaling pathway induced by MDAs (nocodazole, 17α-estradiol, or 2-methoxyestradiol) commonly proceeded through mitotic spindle defect-mediated prometaphase arrest, prolonged Cdk1 activation, and subsequent phosphorylation of Bcl-2, Mcl-1, and Bim in human acute leukemia Jurkat T cells. These microtubule damage-mediated alterations could render the cellular context susceptible to the onset of mitochondria-dependent apoptosis by triggering Bak activation, Δψm loss, and resultant caspase cascade activation. In contrast, when the MDA-induced Bak activation was inhibited by overexpression of anti-apoptotic Bcl-2 family proteins (Bcl-2 or Bcl-xL), the cells in prometaphase arrest failed to induce apoptosis, and instead underwent mitotic slippage and endoreduplication cycle, leading to formation of populations with 8N and 16N DNA content. These data indicate that cellular apoptogenic mechanism is critical for preventing polyploid formation following MDA treatment. Since the formation of polyploid cells, which are genetically unstable, may cause acquisition of therapy resistance and disease relapse, there is a growing interest in developing new combination chemotherapies to prevent polyploidization in tumors after MDA treatment.

돼지고기가 생체내(生體內) 중금속(重金屬)의 축적억제(蓄積抑制)에 미치는 영향

  • Han, Chan-Gyu;Lee, Nam-Hyeong;No, Jeong-Hae;Seong, Gi-Seung;Chae, Chan-Hui
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 1999.06a
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    • pp.97-117
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    • 1999
  • This study was performed to investigate the effect of pork on the cadmium detoxification in rats. Ninety male Sprague-Dawley rats weighing $125.3{\pm}1.4g$ were divided into five groups based on cadmium treatment(0, 25, 50, 100, 250ppm) and five levels of Cd in AIN-76 purified diet had been fed for 8 weeks. Cadmium was supplemented with a form of $CdCl_2$.. During following 8 weeks of intoxication, casein was replaced by pork and the effect of pork on cadmium- detoxification was compared with casein. After 8 weeks of Cd intake had resulted in apparent cadmium intoxication; reduced growth rate, enlarged kidney and testis, decreased hematocrit value and hemoglobin content in response to the supplemented Cd levels in the diets. Discontinuing cadmium feeding, the body weights were relieved. Pork-fed groups seemed to have higher body weight than casein-fed groups. Hemoglobin content and hematocrit value became normal range at detoxification stage. The weights of liver, kidney, and testis were decreased along with cadmium intake. However, organ weight ratio was not affected by cadmium. Cadmium accumulation in liver and kidney showed a tendency to increase in the cadmium-exposed groups. The levels of metallothionein were also significantly elevated in the tissues of liver in response to the levels of Cd supplemented(p<0.05). Cadmium concentration in kidney was two times higher than that in liver. Cadmium removal rate of liver was higher than that of kidney. Cadmium accumulation of the pork-fed group was lower than that of casein. Especially, the factors which affected the cadmium contents in kidney were $Cd^{***}$ and $Cd{\times}pork^{***}$. Metallothionein(MT) was increased with cadmium, and MT was not likely to be affected by pork. Based on the findings from gross lesion, rats fed 250ppm of Cd were externally emaciated, had exposed penis and observed atrophies of kidney and testis. Histopathological observation seemed that the liver of groups feeding Cd supplemented diets showed cellular degeneration and accumulation of eosinophilic materials in the capillaries. In kidney, rats fed Cd diets had shown tubular epithelium degeneration and lesions of basophilic materials, while testis were weakened in numbers of spermatid and sporadically enlarged of giant cells. But the rats administered cadmium-detoxified diet supplemented pork for 7 weeks were shown individually decreased lesions compared with the rats supplied with casein diet.

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Cytotoxic Effects of Prevotella nigrescens on Cultured Cells (Prevotella nigrescens가 배양된 세포에 미치는 영향)

  • Han, Jin-Soon;Kim, Eun-Sook;Lee, Su-Jong;You, Yong-Ouk;Han, Kyung-Soo;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.27 no.2
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    • pp.183-195
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    • 2002
  • 흑색 색소를 형성하는 그람음성 혐기성 세균은 급성 임상 증상을 가진 환자의 근관에서 자주 발견되는 세균으로서 세균 및 세균의 성분과 산물이 치근단 병소의 생성과 밀접하게 연관된 것으로 알려져 있다. 본 연구는 흑색 색소를 형성하는 그람음성 혐기성 세균 중 가장 발현율이 높은 Prevotella nigrescens가 배양된 세포에 미치는 세포 독성을 연구하고자 하였다. 두 가지 세포주 및 사람의 치은섬유모세포를 일차배양하여 사용하였으며, 세포주에 따른 독성 발현에 차이가 있는지를 비교하였다. P. nigrescens ATCC 33563 표준 균주 및 임상 균주로는 환자의 감염된 근관으로부터 165 rRNA primer를 사용한 중합효소 연쇄반응으로 P. nigrescens 6 균주를 동정하여 사용하였다. 세균배양액, 세균의 초음파 추출단백질 및 lipopolysaccharide (LPS)를 MC3T3-El 조골세포, NIH3T3 섬유모세포 및 치은섬유모세포에 첨가한 후 MTT분석법으로 세포의 활성을 측정하였으며, 세포의 형태학적 변화를 도립현미경으로 관찰하였다. 세균배양액을 100$\mu\textrm{l}$ 첨가한 경우는 세가지 세포주 모두에서 통계적으로 유의하게 세포의 활성을 억제하였다. 세균의 초음파 추출단백질 12.5$\mu\textrm{g}$/ml 와 25$\mu\textrm{g}$/ml 는 NIH3T3세포에 통계적으로 유의한 세포독성을 보였다. 세 가지 세포주에 대한 LPS의 세포 독성 효과는 첨가된 LPS의 농도 및 균주에 따라 다양하게 나타났다. 심하게 손상된 세포는 세포의 단일층이 수축되고 세포가 응집되었으며 세포가 배양용기의 바닥에서 떨어지는 양상이 도립 현미경하에서 관찰되었다. 본 연구의 결과 P. nigrescens가 숙주 반응을 조절하여 치수 및 치근단 병소의 유발 및 악화에 기여하는 세균으로 작용할 수 있음을 시사한다. 조직과 함께 제거하고 포르말린에서 48시간 고정시킨 후 파라핀에 포매한 다음에 micro-tome을 사용하여 6$\mu\textrm{m}$로 serial section을 시행하였다. 정중선 부위의 시편에 Hematoxylin-Eosin staining을 시행한 후 Olsson, Orstavik 그리고 Mjor 등의 방법에 따라 조직학적 변화를 관찰한 후 slight(1), moderate(2), severe inflammation(3)의 단계로 분류하였다. 얻어진 결과를 통계처리 프로그램인 Jandel사의 Sigmastat을 이용하여 Kruskal Wallis Test로 통계처리를 하였다. 결과 : (Table omitted) 결론 : 1) Pulp Canal Sealer를 제외한 모든 군에서 시간이 지남에 따라 유의성 있게 염증이 감소되는 양상을 보였다(p<0.05). 2) Pulp Canal Sealer는 1주, 2주, 12주에서 강한 염증반응을 보였다. 3) AH 26과 AH Plus에서는 1주, 2주에서 강한 염증반응을 보였으나 12주에서는 염증반응이 감소하였다. 4) 새로 개발된 봉함제 Adseal-1,2는 1주, 2주에서는 가장 약한 염증반응을 보이나 4주, 12주 후에는 AH Plus와 비슷한 수준의 염증 반응을 보였다. 5) Pulp Canal Sealer를 제외한 모든 군에서 인정할 만한 생체친화성을 보였다. 6) Adseal-2가 Adseal-1에 비하여 전반적으로 낮은 염증반응을 보였다. 7) 각 군간 결과의 차이에 통계적 유의성은 없었다(p>0.05).mmunity. Then, a hierarchical language is to defeat its own purpose.중 행정부가 북한에 대해 실시한 포용정책이 어떠한 성과를 거두고 어떠한 문제점을 간과하고 있는가에 대해 논의하고, 대북 정책의 새로운 지평을 논의하는 것을 목적으로 하고 있다. 1) 포용 정책은 세계의

The Role of Ref-1 in the Differentiation Process of Monocytic THP-1 Cells (단핵구세포주 THP-1의 분화과정에서 Ref-1의 역할)

  • Da Sol Kim;Kang Mi Kim;Koanhoi Kim;Young Chul Park
    • Journal of Life Science
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    • v.34 no.4
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    • pp.271-278
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    • 2024
  • Redox factor (Ref)-1, a ubiquitously expressed protein, acts as a modulator of redox-sensitive tran- scription factors and as an endonuclease in the repair pathway of damaged DNA. However, the function of Ref-1 in the differentiation of monocytes into macrophages has not been defined. In this study, we investigated the effects of Ref-1 on the monocyte differentiation process using the human monocytic cell line THP-1. The differentiation agent PMA increased cell adhesion over time and showed a sig- nificant increase in phagocytic function but decreased the intracellular amount of Ref-1. Ref-1 inhibitor E3330 and Ref-1 knockdown using the siRNA technique reduced cell adhesion and the expression of differentiation markers, such as CD14, ICAM-1, and CD11b, by PMA stimulation. This means that the role of Ref-1 is absolutely necessary in the initial process of differentiating THP-1 cells stimulated by PMA. Next, the distribution of Ref-1 was examined in the cytoplasm and nucleus of THP-1 cells stimulated with PMA. Surprisingly, PMA stimulation resulted in the rapid translocation of Ref-1 to the nucleus. To prove that movement of Ref-1 to the nucleus is required for monocyte differentiation, a Ref-1 vector with the nuclear localization sequence (NLS) deleted was used. As a result, overexpression of ∆NLS Ref-1, which restricted movement to the nucleus, suppressed the expression of differentiation markers and notably reduced phagocytic function in PMA-stimulated THP-1 cells. In conclusion, these data suggest that the differentiation of monocytic THP-1 cells requires Ref-1 nuclear translocation during the initial process of biochemical events following stimulation from PMA.