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发布于:2018-7-12 22:07:30  访问:0 次 回复:0 篇
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Avoid Whining And Initiate Your Own Vemurafenib Venture Preferably
Similar to CO and pectin feeding, YAMC cells treated with LA and butyrate, displayed significantly increased levels of bcl-2 compared to all other groups (Fig. 2b). LA only treatment resulted in a 2-fold increase in bcl-2 expression over control, but LA combined with butyrate induced a 3-fold increase in expression. Furthermore, mRNA analysis using quantitative PCR revealed that LA and butyrate treatment induced bcl-2 transcription (Fig. 2c). These data indicate that the YAMC cell line Vemurafenib mimics in vivo observations. To further probe the contribution of bcl-2 to the effects of DHA or LA when combined with butyrate, we assessed the impact of bcl-2 knockdown on YAMC apoptosis. We hypothesized that decreasing bcl-2 expression in LA and butyrate-treated cells could increase the Venetoclax price level of apoptosis up to that exhibited by DHA and butyrate treatment. YAMC cells were transfected with bcl-2 siRNA, and cultures were harvested 48 hr later. Nontargeting siRNA was used as a control to ensure that any observed changes in apoptosis were not due to off-target effects. Bcl-2 siRNA treatment resulted in a 2-fold decrease in bcl-2 expression compared to untreated control cultures (Fig. 3a). All groups transfected with bcl-2 siRNA displayed at least a 2-fold decrease in bcl-2 expression (Fig. 3b). In contrast, nontargeting siRNA did not significantly alter bcl-2 expression. LA and butyrate-treated cells following siRNA treatment had a comparable level of bcl-2 expression relative to DHA and butyrate-treated cells without siRNA. In subsequent experiments, apoptosis was measured 48 hr post-transfection. Following knockdown of bcl-2, apoptosis in LA see more and butyrate-treated cells was increased to the levels seen in DHA and butyrate cultures (Fig. 4). The proapoptotic effect of DHA and butyrate combination was not influenced by bcl-2 silencing. Since silencing of bcl-2 in LA and butyrate-treated cells elevates apoptosis, we next determined the effects of bcl-2 overexpression. YAMC cells were transfected with full-length human bcl-2 and examined 48 hr later. Transient overexpression subsequently increased bcl-2 levels significantly (Figs. 5a and 5b). Cells treated with either DHA and butyrate or LA and butyrate demonstrated significantly (p < 0.05) inhibited apoptosis when transfected with bcl-2 (Fig. 6). Apoptosis was only inhibited by ?30% in LA and butyrate-treated cells following overexpression of bcl-2. In contrast, overexpression of bcl-2 suppressed apoptosis in DHA and butyrate-treated cells by 2-fold to levels observed in untransfected LA and butyrate-treated cells. The progressive inhibition of apoptosis is closely linked to the transformation of colonic epithelium.3�C5 For this reason, the promotion of apoptosis is an important aspect of chemoprevention.
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