Figure 4

Loss of TARG1 disrupts nucleolar structure and function in U2OS cells. (a) OARD1^−/− and control U2OS cells (KO and WT, respectively) were seeded and fixed in PFA on glass cover slips. A nucleolin antibody was used as to stain nucleoli and was visualised using an AlexaFluor488-coupled secondary antibody. DNA was visualised using Hoechst staining. A representative image of 3 independent experiments is shown. (b) and (c) Quantification of (a) based on automated analysis using pipelines in CellProfiler for nucleolar area and the number of nucleoli per cell, respectively. (d) OARD1^−/− and control U2OS cells were seeded in 8-well Ibidi slides and fixed in PFA. Transcription rates were measured using a CLICK-iT kit (EdU) and (e) quantified as a median intensity per nucleus using CellProfiler software. Scale bars represent 10 µM. At least 200 cells were measured for quantification of nucleolus number/footprint and transcription rate presented in b and e, respectively. Statistical analysis was performed by a Wilcoxon rank-sum test and corresponding p-values are shown in plots.