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Own MR, Speirs V: Immunohistochemical detection of ERbeta in breast cancer: towards more detailed receptor profiling? Br J Cancer 2001, 84(8):1095?098. 33. Zhao C, Dahlman-Wright K, Gustafsson JA: Estrogen receptor beta: an overview and update. Nucl Recept Signal 2008, 6:e003. 34. Saji S, Hirose M, Toi M: Clinical significance of estrogen receptor beta in breast cancer. Cancer Chemother Pharmacol
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Collected and analyzed for de novo synthesis and activity of MMPs by gelatin zymography. As shown in Figure 1A, pretreatment with TSIIA (0.110 M) significantly attenuated LTA-induced proMMP9 expression and activity. Moreover, pretreatment with TSIIA (10 M) also markedly inhibited LTA (50 g/ml, 16 h)-induced MMP-9 mRNA expression, determined by RT-PCR (Figure 1B), suggesting that AP-1 is an importa
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Lected and disrupted by sonication in lysis buffer (25 mM Tris, pH 7.8, 2 mM EDTA, 1 Triton X-100, and 10 glycerol). After centrifugation, aliquots of the supernatants were tested for luciferase activity using the luciferase assay system. Firefly luciferase activities were standardized for b-galactosidase activity.To detect the in vivo association of nuclear proteins with rat mmp-9 promoter, chr
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Yrophosphate, 1 sodium vanadate, 2.5 EDTA, 2.5 EGTA, 0.05 (w/v) Triton X-100, 0.5 (w/v) SDS, 0.5 (w/v) deoxycholate, 0.5 (w/v) NP-40, 5 mg/ml leupeptin, 5 mg/ml aprotinin, and 1 PMSF. The lysates were centrifuged at 45,000 ?g for 1 h at 4 to yield the whole cell extract. The protein concentration was determined by the BCA reagents according to the instructions of the manufacturer. Samples fro
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Ted and analyzed by RT-PCR. (C) Time dependence of LTA-stimulated JNK phosphorylation. RBA-1 cells were pretreated with SP600125 for 1 h and then treated with 50 mg/ml LTA for the indicated time intervals. Phosphorylation of JNK and c-Jun was determined by western blot using an anti-phospho-JNK or phospho-c-Jun antibody. (D) Cells were transfected with an empty vector (pcDNA3, as a control) or dom
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Was added to the cells. The figure represents one of at least five similar experiments. (F) Effects of calcium inhibitors on LTA-induced phosphorylation of JNK and c-Jun, RBA-1 cells were pretreated with BAPTA or TG for 1 h and then incubated with 50 mg/ml LTA for the indicated time intervals. Phosphorylation of JNK and c-Jun was determined by western blot using an anti-phospho-JNK or phospho-c-Ju
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Were covered with 1 ml of DMEM/F-12 containing 5 M Fura2/AM and incubated at 37 for 45 min. At the end of the period, the cover slips were washed twice with the physiological buffer solution containing (in mM): 125 NaCl, 5 KCl, 1.8 CaCl2, 2 MgCl2, 0.5 NaH2PO4, 5 NaHCO3, 10 HEPES, and 10 glucose, pH 7.4. The cells were incubated in physiological buffer for further 30 min to complete dye de-esterif
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Own MR, Speirs V: Immunohistochemical detection of ERbeta in breast cancer: towards more detailed receptor profiling? Br J Cancer 2001, 84(8):1095?098. 33. Zhao C, Dahlman-Wright K, Gustafsson JA: Estrogen receptor beta: an overview and update. Nucl Recept Signal 2008, 6:e003. 34. Saji S, Hirose M, Toi M: Clinical significance of estrogen receptor beta in breast cancer. Cancer Chemother Pharmacol