Correction: Misregulation of AUXIN RESPONSE FACTOR 8 Underlies the Developmental Abnormalities Caused by Three Distinct Viral Silencing Suppressors in Arabidopsis
Fig 5
RNAi and miRNA-mediated gene silencing are not compromised by the arf8 mutation.
(A) Western analysis of DCL1 accumulation in arf8 homozygous seedlings compared to WT seedlings and AGO1 accumulation in arf8 homozygous flowers compared to WT flowers. Due to the strong developmental defects (dcl1) or developmental arrest (ago1) exhibited by miRNA deficient mutants, the negative controls used in this analysis were from seedlings with the hypomorphic dcl1–9 genotype, which accumulate a truncated, non-functional form of the DCL1 protein, and from seedlings with the null ago1–36 genotype. Load: Coomassie staining provides a control for equal loading of total proteins. (B) Northern analysis of various endogenous miRNAs in Col-0 or homozygous arf8 mutant seedlings. (C) qRT-PCR analysis of transcript levels from various targets of the miRNAs studied in (B), showing intact miRNA-mediated repression in arf8 mutants as compared to WT plants. (D) RNAi of CHS, diagnosed by a loss-of-seed pigmentation (inlays), remains unaltered in plants with the arf8-/- genotype. (E) Northern analysis of CHS siRNAs and endogenous miRNA accumulation in VSR transgenics with the heterozygous arf8 mutant background (as depicted in Fig 5B-I). Note the strong decrease in CHS siRNA levels caused by HcPro and P15 as well as the slight shift in electrophoretic mobility and increased accumulation incurred to miRNAs by P19 and HcPro, respectively. The inlays at the bottom show that RNAi of CHS remains suppressed by the three VSRs in the arf8 mutant background, as diagnosed by the dark-brown seed coloration. (F) qRT-PCR analysis of transcript levels from various targets of the miRNAs studied in (B) in the P19 transgenics carrying the homozygous arf8 mutation (CHS RNAi background), as depicted in Fig 5H. Reference plants used in the analysis were line CHS RNAi and its P19 transgenic derivative (P19 CHS RNAi) with a wild type background. Off-scale values for ARF17 and At4g22470 (a novel small RNA target shown in Fig 4A) are indicated by double-dashed lines. U6: oligonucleotide hybridization of the ubiquitous U6 small nucleolar RNA provides a control for equal RNA loading.