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. Author manuscript; available in PMC: 2009 Nov 1.
Published in final edited form as: Nat Genet. 2009 Apr 19;41(5):630–634. doi: 10.1038/ng.365

Figure 4. AGO4 physically interacts with Pol V transcripts.

Figure 4

(A) RNA immunoprecipitation using αAGO4 antibody. Immunoprecipitated RNA isolated from the indicated mutants was digested with DNaseI and amplified by RT-PCR. Total RNA controls show that the Pol V transcripts are present in equivalent amounts in all mutants tested except nrpe1. Ethidium bromide stained rRNAs (bottom left) show that equal amounts of RNA were tested. The no reverse transcriptase (no RT) control was performed with IGN5 bottom strand primers. No RNA controls were performed for all primer pairs tested. RT-PCR amplification of actin RNA serves as a loading control.

(B) Immunoblot detection of AGO4 in protein extracts of wild-type (Col-0) plants or ago4 mutant. Stars denote nonspecific bands.

(C) Immunoblot detection of AGO4 in protein extracts of wild-type (Col-0), rdr2, dcl3, dcl234 or nrpe1 mutants. Asterisks denote nonspecific bands.

(D) Immunoblot detection of AGO4 in protein extracts of wild-type (Col-0), nrpd1 (Pol IV) nrpe1 (Pol V), nrpd2/nrpe2 (shared subunit of Pol IV and Pol V), or rdr2 mutants. Asterisks denote nonspecific bands.