Percentage yield of thiol-labeled RNA from hippocampal neurons after subcutaneous administration of 4TU
| Sample type | Age | Tissue | Percent yield |
|---|---|---|---|
| TetO-UPRT; CaMKII-tTA | P50 | Hippocampus | 3.90% |
| TetO-UPRT; CaMKII-tTA | P90 | Hippocampus | 3.20% |
| TetO-UPRT | P120 | Hippocampus | 2.00% |
| TetO-UPRT | P120 | Hippocampus | 2.00% |
| WT | P50 | Hippocampus | 0.07% |
| WT | P90 | Hippocampus | 0.10% |
CaMKII-tTA; TetO-UPRT double positive mice, TetO-UPRT single positive mice, or WT control mice were injected subcutaneously with 4TU and hippocampus was removed 5 h later. After RNA extraction and streptavidin purification, the amount of tagged RNA obtained was much higher from hippocampal samples expressing UPRT in neurons (CaMKII-tTA driven neuronal expression and leaky neuronal expression in single positive TetO-UPRT mice; Fig. 6-2).