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. 2009 Nov 11;38(2):360–369. doi: 10.1093/nar/gkp977

Figure 2.

Figure 2.

The 7SK-independent interaction between MePCE and LARP7 nucleates the formation of a stable MePCE–LARP7–7SK subcomplex within 7SK snRNP. (A) NEs prepared from W10, a HeLa-based cell line stably expressing F-MePCE, and the parental HeLa cells were analyzed by WB for the indicated proteins. (B) W10 cells were treated with the indicated agents. NEs prepared from the treated cells were analyzed by WB with the indicated antibodies (right panel) and subjected to anti-Flag immunoprecipitation. Upon elution with the Flag peptide, the compositions of the immunoprecipitates (αFlag IP) were analyzed by WB and NB as indicated (left panel). (C) Prior to the elution with the Flag peptide, the immobilized αFlag IP were washed with a buffer containing the indicated KCl concentrations, and their compositions were subsequently analyzed as in B. (D) NEs of W10 (left panel) and FPS86 cells, a HeLa-based cell line stably expressing F-LARP7 (right panel), were incubated with (+) or without (−) RNase A prior to anti-Flag immunoprecipitation. αFlag IP were analyzed as in B. (E) NEs of HeLa cells were pretreated with (+) or without (−) RNase A prior to immunoprecipitations with the indicated antibodies. The immunoprecipitates were analyzed as in B.