The effect of PAX3 and MITF knock-down in melanoma cell lines
siRNA-mediated knockdown of PAX3 and MITF in NZM12 and NZM15
- Reverse transfection with Dharmacon Smartpools (10 nM MITF; 20 nM PAX3; 20 nM non-targeting pool).
- Assess transcript knock-down at 24 hours post-txn by using qPCR.
- Assess protein levels 48 hours post-txn by using Western.
- n = 2 of 3 replicate experiments.
- Measure EDNRB, RAB27A, FOXD3, and SNAI2 transcript levels by using qPCR at 24 and 48 hours post-txn.
Does miR-137 repeat expansion correlate with MITF levels in NZM cells?
- Assess miR-137 locus stability at 1p22 with conventional PCR.
- purify and sequence a selection of the different sized amplicons.
- predict impact of repeat expansion on pre-miR folding.
- measure miR-137 VNTR status in non-melanoma/skin-derived cells?