Fig. 7.

6T/S-A NeuroD4 shows enhanced protein stability and enhanced chromatin binding relative to WT NeuroD4 (a-d). Embryos were injected at the one cell stage with 200 pg mRNA encoding HA-tagged versions of either WT or 6T/S-A NeuroD4. a Western blot analysis on extracts prepared from stage 13 embryos, with tubulin as a loading control. b The density of the protein band for WT or 6T/S-A NeuroD4 was quantified and expressed relative to the tubulin loading control of each sample. Mean values are shown from independent duplicate samples with the standard error of the mean. c At stage 14, cross-linking and chromatin isolation were performed prior to western blot analysis as before. No HA-tagged NeuroD4 protein was detected in the uninjected embryos confirming antibody specificity, and no tubulin protein was detected in the chromatin fraction. d NeuroD4 protein bands were again quantified relative to tubulin or histone H3 bands for cytoplasmic and chromatin fractions respectively. Mean values are shown from independent duplicate samples with the standard error of the mean