Fig. 1: The dual reporter tethering assay reports reproducible and quantitative regulatory effects.
From: Surveying the global landscape of post-transcriptional regulators
a, Schematic of the tethering assay with a YFP reporter and RFP control (top), with expected fluorescence levels based on the activity of the tethered query protein (bottom). b, Schematic of testing the effects of the reporter mRNA and protein-RNA tether. c, The fluorescence ratio changes from tethering Pab1 and Pop2 to fluorescent protein reporter mRNAs, relative to tethering an inactive Halo control. Multiple regression (adjusted R2 = 0.996, F(7, 8) = 563.2, P = 3.9 × 10−10) indicated significant effects of Pab1 (log2 change of 1.35, 95% CI 1.18–1.53, t = 17.8, P < 0.001), Pop2 (log2 change of −2.40, 95% CI −2.58 to −2.23, t = −31.7, P < 0.001) and tether choice on Pop2 (log2 change of 0.92, 95% CI 0.57–1.26, t = 6.0, P < 0.001); no other terms were significant (all P values are two-sided with no multiple testing correction). d, Distribution of fluorescence ratios reporting on the activity of Sgn1 in the tethering assay in two replicate samples. The dashed line represents the median YFP expression.
