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. 2015 Jun 15;5(1):10–24.

Figure 6.

Figure 6

AF4N/AF4 transfers P-TEFb from 7SK RNPs to AF4N/AF4. 293T cells were transiently transfected with empty vector (mock), pEZP-AF4ST or pEZP-AF4NST. Whole cell lysates were prepared and separated on a 0-40% glycerol gradient by ultracentrifugation. Gradient were fractionated (n = 10) and analyzed for the presence of CDK9, LARP7 and AF4/AF4N. Dissociation of LARP7 from 7SK RNPs is equivalent to the disruption of existing 7SK RNPs. Bottom: AF4 immunoprecipitation from glycerol gradient fractions 2 to 7 was probed for the presence of CDK9 to demonstrate the P-TEFb transition from 7SK RNP to AF4N/AF4.