The experiment involved the use of 368T1 murine lung cancer cells expressing either an empty vector (empty), a vector expressing wild-type Hmga2 (wt), a vector expressing a let-7 site mutated Hmga2 (m7), a vector expressing wild-type Hmga2 without a start codon (ATG wt), or a vector expressing a let-7 site mutated Hmga2 without a start codon (ATG m7).
I used the Millipore MAGNA-RIP kit with 5 ug Ago2 or IgG antibody per 1/2 of a 15 cm plate of cells (approximately 10^7 cells each). Following the RIPs, I did qRT-PCR using the Qiagen miScript system to detect miR-10a associated with IgG/Ago2 in these cells.
Overall, we observed no difference in miR-10a association between the samples but a clear enrichment (note the log scale) of miR-10a in the Ago2 IPs versus IgG.
Abcam user community
Submitted Nov 15 2012
Get resources and offers direct to your inboxSign up