Figure 4. Characterization of Bcl-xL effects on NF-κB activity.A: Fertilized eggs were injected with RNA (650 pg/embryo) encoding Xenopus IκBα-V5. Beginning at stage 8, experimental embryos were treated with 50 µM AKBA and analyzed at stage 11 by SDS-PAGE/immunoblot using an anti-V5 antibody and the antiSOX3c antibody to visualize endogenous Sox3 protein as a loading control. AKBA treatment stabilized the IκBα-V5 polypeptide. B: Fertilized eggs were injected with p3XκB-firefly luciferase (“3κB-Luc”) and pTK-Renilla luciferase (“RL-TK”) DNAs (10 pg/embryo each) either alone (“Con”) or together with Bcl-xL (500 pg/embryo) RNA, or Bcl-xL and IκBsa (600 pg/embryo) RNAs. Alternatively, animal caps prepared from Bcl-xL RNA injected embryos were cultured in either control buffer (0.1% DMSO), 20 µM or 50 µM AKBA. At stage 11, caps were analyzed for luciferase activity. Bcl-xL induced an increase in 3XκB-Luc activity that was blocked by either IκBsa or AKBA. Error bars in B reflect standard deviation from the mean of multiple experiments.
Image published in: Zhang C et al. (2006)
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