Mapping of MLE and MSL2 using iCLIP
Mapping of MLE and MSL2 using iCLIP
Clone 8 cells, expressing roX1 and roX2, were grown in M3 + 2% FCS + 5 ug/ml insulin + 2.5% fly extract at 25C. We have isolated nuclei, and used nuclear extracts in all experiments. We used Clone8 cells and antibodies against MLE (rat1) and MSL2 (d300, Santa Cruz). One 150mm dish of ~80% confluent cells is used per immunoprecipitation; 6uL from each antibody and 100uL slurry of Protein G (for MLE) or protein A (MSL2) Dynal beads (Invitrogen). The low-RNaseI (Ambion) concentration used for library-preparation was 0.5U/mL. Biological triplicates were tagged with different barcodes and mixed together after reverse-transcription.
Experimental Factor: immunoprecipitate
MLE antibody (rat1)