From: Pax7 is back
TMX delivery | Recovery period | Gene deletion analysis | Escaper cell analysis |
---|---|---|---|
TMX injection at 3 mg/day for 5 consecutive daysa | Allow >15 days of TMX chase prior to muscle injury to target quiescent SCs and minimize lingering TMX activity | Isolate purified SCs by FACS, or from single fibers from each experimental animalb | Measure gene or protein levels over timec |
Five daily injections prior to injury followed by continuous TMX feeding delivered in chow (1 mg TMX per day) | Pros: Temporal identity of cell type and state | Measurement of protein by western blot or immunohistochemistry | In vitro: Measure gene/protein immediately after isolation and after approximately 10 days in culture |
Minimize recombination of transient cell populations | Quantify DNA directly at the gene locus using site-specific primers flanking the loxP sites | In vivo: Measure gene/protein from SCs isolated from uninjured and regenerated muscle (>30 days after injury) | |
Cons: Escaper cell contribution | |||
No recovery period | |||
Pros: Maximize targeting of both stable and transient cell populations | |||
Minimize escaper cell contribution | |||
Cons: Lose cell type and state resolution |