Figure 4. Placement of SR45 in RdDM pathway. (A) RNA gel blots showing siRNAs abundance at both type I and II loci. Hybridization with miR163 is shown as a loading control for 5S siRNAs and hybridization with miR159 is shown as a loading control for AtSN1 and siR02. (B) RNA gel blot showing AGO4 expression in leaves, seedlings, and flowers. Hybridization with UBQ10 is shown as a loading control. (C) RT-PCR and protein gel blot showing the expression and abundance of AGO4/AGO4. UBQ10 expression is showed as a loading control for RT-PCR and amido black staining of the RUBISCO large subunit is shown as a loading control for the protein gel blot. (D) Immunofluorescent microscopy showing AGO4 localization in 4xmyc::AGO4 (Columbia) and 4xmyc::AGO4 (sr45–1) backgrounds. White arrows indicate the position of AGO4 in the sr45–1 panel.