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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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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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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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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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With the cells exposed to LTA alone. The figure represents one of three individual experiments.TG-sensitive Ca 2+ stores and the extracellular Ca 2+ influx which is essential for LTA-induced proMMP-9 expression in RBA-1 cells. To further determine the effect of Ca2+ signaling on LTA-stimulated JNK/c-Jun cascade, the JNK and c-Jun phosphorylation stimulated by LTA in the presence of BAPTA/AM or TG
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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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De and up-regulation of MMP-9 by LTA. A recent study has indicated that induction of MMP-9 by IL-1b is mediated through Ca 2+ -dependent signaling [35]. Hence, we investigated the role of intracellular Ca 2+ in LTAinduced proMMP-9 expression, the intracellular Ca2+ chelator BAPTA/AM and ER Ca 2+ -ATPase blocker thapsigargin (TG) were used. As shown in Figures 3A and 3B, pretreatment with BAPTA/AM
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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