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. Author manuscript; available in PMC: 2012 Oct 1.
Published in final edited form as: Biosci Rep. 2011 Oct 1;31(5):333–343. doi: 10.1042/BSR20100124

Figure 6. The MeCP2-Prpf3 complex interacts with mRNA in vivo.

Figure 6

(A) RT-PCR analysis of a RIP from HA-MeCP2 stably transfected HT-22 cells indicates an association of HA-MeCP2 with Cdk10 mRNA (top, lane 2), and FRG1 mRNA (middle, lane 2), but not Casc3 mRNA (bottom, lane 2). Control RIPs using normal rabbit serum (IgG) (lane 3) show no RT-PCR product. RT-PCRs using unbound RNA from RIPs indicate target mRNAs were present in all RIP samples (lanes 4 and 5) and all RT-PCRs were RNase sensitive (+ RNase) confirming RNA and not DNA was being assayed. (B) An anti-HA RIP followed by anti-Prpf3 Re-RIP experiment from HA-MeCP2 HT-22 cells was assayed for Cdk10 mRNA by RT-PCR (lane 2). Control RIP and Re-RIP experiments using normal rabbit serum (IgG) showed no product by RT-PCR (lane 3). Unbound mRNA was assayed by RT-PCR for the RIP (lanes 6 and 7) and Re-RIPs (lanes 4 and 5) to confirm the presence of the Cdk10 mRNA in the reactions. All RT-PCRs were sensitive to RNase treatment (lower panel) confirming the amplifications were from RNA and not DNA templates.