Changes in Acetyl CoA Levels during the Early Embryonic Development of Xenopus laevis
Figure 6
Microinjection of acetyl CoA to stage one embryo increases protein acetylation levels.
Immediately after dejellying (30–45 minutes after fertilisation), stage one embryos were microinjected with increasing amounts of acetyl CoA, as described in the Materials and Methods section. Control embryos were injected with water. Injected embryos were allowed to progress to stage 3 and collected for HPLC analysis of CoA compounds or Western blot analysis with anti-acetyl-lysine antibody. For both analyses, 10 injected embryos were used per condition. In addition, embryos were collected after reaching stage 9. Microinjection of acetyl CoA dose dependently increased acetylation levels of several proteins (labelled 1–4). Equal protein loading was confirmed by Ponceau staining of the blot. A section of the Ponceau stained membrane is shown (the complete membrane is shown in Figure S4). For the Western blot shown, linear contrast and brightness adjustment was applied uniformly to the whole blot for clarity. The intensity/size of each of the bands numbered 1–4 was quantified and expressed relative to control embryos.
