Labeling of nascent RNA in the C. elegans intestine
Fig 2
EU-click can measure changes in nascent RNA transcription in the C. elegans intestine.
Images and quantitative measurements of nascent transcription (5 min pulse) with and without transcription inhibition. (A) Example nucleoli from int1 cells after treatment with DMSO (top) or ActD (bottom). Nucleolar vacuole is marked (*). Columns from left to right: GARR-1::GFP (FZ marker), EU conjugated with AF594 (EU-AF594), composite image of GARR-1::GFP (green) and EU-AF594 (orange), and intensity profiles of GFP and EU-AF594 signal along the line shown on the composite image. EU-AF594 pixel values in the grayscale and composite images and the line plots are normalized using the same range for DMSO and ActD. (B) Transcription rate (EU incorporated/5 min) in the nucleolus decreases upon transcription inhibition by ActD. Box plot comparing the sum of pixel values in the nucleolar mask for DMSO- and ActD-treated int1 cells. Points represent measurements from different worms; color indicates batch number. Significance was tested by two-way ANOVA using batch number and treatment as independent variables (p < 1 × 10-8). (C) Example nucleoli from int1 cells from L4440 (control) and tif-1a KD. Similar to A. (D) Transcription rate in nucleolus decreases upon tif-1a KD. Measurements and test of significance are similar to B (p < 1 × 10-14).