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. 2021 Aug 24;12:5107. doi: 10.1038/s41467-021-25433-6

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© The Author(s) 2021, corrected publication 2022

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Fig. 7 — A Electrophoretic mobility shift assays show the in vitro binding for the wild-type PUF domain, the QFR/VFQ PUF variant, and the design 2 variant to the wild-type RNA sequence or an RNA containing mutated bases. Results are representative of two biological replicates. B Yeast three-hybrid assays show the binding of the wild-type PUF domain and QFR/VFQ variant. The negative control is a wild-type PUF domain that has stop codons in the TRM locations (repeat 1) paired with wild-type RNA. SC-L, synthetic complete media minus leucine; SC-L-H, synthetic complete media minus leucine and histidine. C Spot dilution plate assay indicates the binding of the design 1 and design 2 PUF domains to their target RNAs. The negative control is as in (B). The starting OD600 that was spotted was 0.05, with three sequential 10-fold serial dilutions shown.