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. 2023 May 23;299(7):104859. doi: 10.1016/j.jbc.2023.104859

Figure 2.

Figure 2

In vitro transcription and PIC formation analyses.A, a representative autoradiograph showing SUP4 tRNA transcripts from a multi-round transcription assay using whole cell extracts (WCE) from indicated yeast strains. Mutations of Rpc53 are indicated. B, scatter plot reporting the reproducibility of independent biological replicates shown with filled circles (n = 3). Error bars represent the mean ± SD values from three independent experiments using separately prepared WCEs. Unpaired t test was used to detect significant differences compared with each sample to the WT (Wild Type) (∗∗∗∗ denotes p < 0.0001, ∗∗∗ denotes p ＝ 0.0001, and ∗∗ denotes p = 0.003). C, western analysis of proteins in the isolated pol III PICs from the immobilized template assay. The DNA template contained the SUP4 tRNA gene. The positions of the molecular weight markers (kDA) are shown between the Western blotting panels. The gel bands corresponding to each indicated protein subunit were validated by specific antisera for individual polypeptides.