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. 2018 Oct 11;30(11):2838–2854. doi: 10.1105/tpc.18.00244

Figure 3.

Figure 3.

Arabidopsis LUC7 Is an U1 snRNP Component.

(A) RNA immunoprecipitation using a pLUC7A:LUC7A-YFP luc7a luc7b luc7rl complementation line. Proteins were immunoprecipitated using GFP-affinity matrix and RNAs were extracted from the input and the immunoprecipitated fraction. U1, U2, U3 snRNAs, and ACTIN RNA were quantified using RT-qPCR. Enrichment of the respective RNA in pLUC7A:LUC7A-YFP luc7a luc7b luc7rl transgenic line was calculated compared with the wild type (negative control). Error bars denote the range of two biological replicates.

(B) Quantification of U1 and U2 snRNA levels in the wild type and luc7a luc7b luc7rl mutants. Total RNAs were isolated from 7-d-old seedlings, and RNA levels were analyzed by RT-qPCR using U1- or U2-specific oligonucleotides. Error bars denote the range of two biological replicates.

(C) Subcellular localization of LUC7A in pLUC7A:LUC7A-YFP luc7a luc7b luc7rl in Arabidopsis transgenic plants. Roots of 9-d-old seedlings were analyzed using confocal microscopy. Bar = 25 µm.

(D) U1-70K-mRFP and LUC7A-YFP or LUC7RL-YFP proteins were transiently expressed in N. benthamiana and their subcellular localization analyzed using confocal microscopy. Bars = 10 μm and 25 µm for upper and lower panels, respectively.