Figure 5.

RPAP2 Is Required for Proper Expression of Pol II-Transcribed snRNA Genes

(A) RNase protection analysis of transcripts from endogenous U2 snRNA genes in control cells or cells transfected with an siRNA specific for RPAP2 (left panel). The different RNA species are noted.

(B) qRT-PCR analysis of U2, pre-U2, and RPAP2 mRNA in total RNA normalized to the Pol III-transcribed 7SK RNA.

(C) qPCR quantitation of ChIP analysis of U2 snRNA genes using antibodies against the Int5 and Int11 subunits of the Integrator complex, after RPAP2 knockdown. The regions amplified are noted on the diagram above.

(D) Nuclear run-on analysis with and without RNAi-mediated knockdown (KD) of RPAP2 is shown. An oligonucleotide complementary to transcripts from the Pol III-transcribed 5S RNA gene was used as a control for the level of transcription. Corrected hybridization signals are shown in the graph below.

(E) qPCR quantitation of ChIP analysis of U2 snRNA genes using an antibody against Pol II occupancy, with or without RPAP2 knockdown. Regions amplified are as shown in (C).

Error bars indicate the standard deviation obtained from at least three independent experiments. See also Figure S5.