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Genetic Analysis of NRPD1 and AGO4 Mutants Reveals Insights into RNA Silencing Mechanisms

This study examines the impact of various genetic mutations, including nrpd1a, nrpd1b, and ago4, on RNA silencing and gene expression regulation in Arabidopsis. Utilizing Southern blot analysis and other molecular techniques, we investigate the deletion variants and their effects on marker gene expression under different temperature conditions. Our results highlight the roles of RDM proteins in controlling RNA-dependent DNA methylation and the expression of stress response genes, contributing to a deeper understanding of plant RNA silencing pathways.

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Genetic Analysis of NRPD1 and AGO4 Mutants Reveals Insights into RNA Silencing Mechanisms

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  1. He_Supp. Fig.1 ( ) rdm5 nrpd1a ATG (1) TGA (4362) ( rdm6 - 2 nrpd1b) ( ) rdm6 - 1 nrpd1b 1377 bp deletion ATG (1) TAA (5931) ( ) rdm8 ago4 6 bp deletion ATG (1) TAA (2775) ( ) rdm9 hen1 TGA (2829) ATG (1) rdm10 (hda6) 13 bp deletion TAA (1416) ATG (1) rdm11 (drd1) 35 bp deletion TAA (2667) ATG (1) A AT1G63020 B AT2G40030 C AT3G23780 D AT2G27040 E AT4G20910 F AT5G63110 G AT2G16390

  2. He_Supp. Fig.2 A nrpd1b B ago4 ros1 ros1 ros1rdm6-1 ros1rdm8 WT WT 23 °C 23 °C 23 °C 23 °C 23 °C 23 °C 4 °C 4 °C 4 °C 4 °C 4 °C 4 °C RD29A RD29A LUC NPTII NPTII COR15A COR15A TUB8 TUB8 rRNA rRNA C hda6 D drd1 ros1 ros1rdm10 WT ros1 ros1rdm11 WT 23 °C 23 °C 23 °C 4 °C 4 °C 4 °C 23 °C 23 °C 23 °C 4 °C 4 °C 4 °C RD29A RD29A LUC LUC NPTII NPTII COR15A COR15A TUB8 TUB8 rRNA rRNA

  3. He_Supp. Fig.3 A 5S rDNA repeat Southern ros1nrpd1b ros1nrpd1b ros1nrpd1b ros1hda6 ros1ago4 ros1hda6 ros1ago4 ros1hda6 ros1ago4 ros1drd1 ros1drd1 ros1drd1 ros1 ros1 ros1 WT WT WT HpaII MspI HaeIII B ros1nrpd1b ros1hda6 ros1ago4 ros1drd1 ros1 ros1 ros1 ros1 WT WT WT WT AtSN1 HaeIII AtSN1 No Digest D C ros1ago4 ros1hda6 ros1drd1 ros1nrpd1b ros1hda6 ros1ago4 ros1drd1 ros1 ros1 ros1 WT WT WT ros1 WT AtGP1 siRNA1003 microRNA173 AtMU1 U6 snRNA TUB8

  4. He_Supp. Fig.4 nrpd4-2 nrpd1a nrpd1b Col-0 Cluster4 siRNA AtSN1 siRNA siRNA1003 AtGP1 siRNA AtMU1 siRNA siRNA02 siRNA255 microRNA171 tRNA and 5S rRNA A B 180 bp CEN nrpd4-2 nrpd4-2 nrpd4-2 nrpd1a nrpd1b nrpd1a nrpd1b nrpd1a nrpd1b Col-0 Col-0 Col-0 HpaII MspI HaeIII

  5. He_Supp. Fig.5 B A SD-Trp-Leu-His +50 mM 3AT SD-Trp-Leu C

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