Fig. 5: The tethering screen identifies RNA-regulatory roles of poorly characterized proteins.
From: Surveying the global landscape of post-transcriptional regulators
a, Schematic representation of the Gta1 protein, with the C-terminal Gta1(603–767) fragment highlighted. b, Schematic depiction of Gta1(603–767) in the tethering assay. c, Flow cytometry measuring activity of Gta1 and Gta1(603–767) in the tethering assay, where dashed lines represent the median YFP expression (n = 2, one replicate per sample is shown). d, RT–qPCR analysis of YFP mRNA abundance with Gta1 tethered to the 3′ UTR, normalized to a non-regulator control (n = 3 independent biological replicates are shown in different shades; P = 0.00026, two-sample t-test with unequal variance). e–g, Time course of reporter changes after induction of Gta1 (e), Gta1Δ603–767 (f) and Halo control tethering constructs (n = 2) (g). h, Change in BFP fluorescence as a measure of Gta1 expression over time, normalized to the uninduced Gta1 sample (n = 2, one replicate is shown per time point). i, As in h, for Gta1Δ603–767. j, RT–qPCR analysis of induced GTA1 relative to endogenous GTA1 expression (n = 3 biological replicate cultures). k, Light microscopy images of yeast overexpressing GTA1, GTA1Δ603–767 or Halo control for 4 h (n = 3 biological replicate cultures are shown, with two frames per replicate). Scale bar, 20 µm.
