Extended Data Fig. 6: The GHR motif is crucial for miRNA expression and function.
From: Key RNA elements influencing DCL1 cleavage in plant microRNA biogenesis
(a) Structural diagrams of various WT and mutant pre-miR172a with high or low GHR scores. The arrowheads indicate the cleavage sites of DCL1. Red letters indicate the mutant nucleotides. (b) In vitro DCL1 cleavage assays for pre-miRNAs with varying GHR scores. The blue and black arrowheads indicate the F1/3 and F2 fragments of DC21, respectively. (c) Quantitative analysis of relative cleavage efficiency for three high-GHR and three low-GHR pre-miRNAs compared to WT pre-miR172a, based on data from Fig. 6b and (b). Bars represent the relative cleavage efficiency of each mutant pre-miR172a normalized to pre-miR172a WT, data values for each pre-miRNAs are shown as dots and error bars represent mean ± SD. The p-values were calculated by a two-tailed t-test. (d) In vitro DCL1 cleavage assays for pre-miR172a variants with DCL1ΔdsRBD1. The blue and black arrowheads indicate the F1/3 and F2 fragments of DC21, respectively. The assays were repeated two times with similar results. (e) The relative expression of miR172a/b/e by qPCR in individual lines. (f) The relative expression of two genes, TOE1 and SMZ, by qPCR in individual lines. For (e) and (f), the number of plants (n = 10/8) for each line is indicated below the corresponding boxplot, the center line represents the median, the box’s lower and upper bounds correspond to the 25th and 75th percentiles, whiskers show 1.5x the interquartile extending from bounds of box, minima is the minimum value, and maxima is the maximum value. The p-values were calculated by a two-tailed t-test.
