Fig. 3: Dbp2 is a component of cleavage bodies. | Nature Communications

Fig. 3: Dbp2 is a component of cleavage bodies.

From: DEAD-box ATPase Dbp2 is the key enzyme in an mRNP assembly checkpoint at the 3’-end of genes and involved in the recycling of cleavage factors

Fig. 3

A Live cell imaging of GFP-tagged RNA processing factors and the MTREC component Red1-tdTomato as a marker for cleavage bodies. The merged channel shows GFP in green and Red1-tdTomato in magenta. The sketch below indicates the typical localisation observed for the GFP-tagged protein in at least three independent experiments. B Live cell imaging of genomically tagged Dbp2-GFP and the nuclear poly(A)-binding protein Pab2-tdTomato. Images are representative of three independent experiments. Fluorescence intensity profiles were generated along the yellow line. C Fluorescence intensity profiles along the lines indicated in B. Green line corresponds to the Dbp2-GFP signal, dashed magenta line to the Pab2-tdTomato signal. Fluorescence intensities were normalised to a 0-1 range. Source data are provided as a Source Data file. D Live cell imaging of mixed cultures of wild type (marked by NLS-BFP; blue outlines) and an analogue-sensitive mutant of Cdk9 (cdk9-as; yellow outlines) with either Pcf11 or Rna15 tagged with GFP. Cells were grown in in YES at 30 °C and treated with 100 µM 3-MB-PP1 for 10 min to block transcription elongation before pelleting and resuspension in EMMG + 3-MB-PP1 for imaging. The GFP channel is shown as the maximal intensity projection using a blue orange icb colour scale as indicated. Images for the untreated controls are provided in Suppl. Fig. 3D, E. E Quantification of the inhibitor experiment shown in (D) (n = 2 independent experiments). Measurements were performed on maximal intensity Z-projections and the maximal fluorescence intensity scored for each cell. Non-treated mixed cultures were included as controls. The numbers below the graph indicate the total number of counted cells for each condition. The lower and upper hinges correspond to the first and third quartiles, and the whiskers extend from the hinge to the smallest and largest value no further than 1.5 * IQR from the hinge (where IQR is the inter-quartile range). The horizontal line marks the median value. The displayed p-values for the pair-wise comparisons were calculated using a two-sided Wilcoxon rank sum test. Source data are provided as a Source Data file.

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