Fig. 5
Poly-GR proteins form cytoplasmic inclusions in HEK-293 T cultured cells. a Expression of GFP-(GR)50 in HEK-293 T cells accumulate in nucleus. No cytoplasmic inclusions are detected. Expression of GFP-(GR)100 in HEK-293 T cells show immunoreactivity in nucleus and cytoplasm, with formation of cytoplasmic inclusions. Double immunofluorescence of GFP-(GR)100 and aDMA in HEK-293 T cells. b GFP-(GR)100 and aDMA. GFP-(GR)100 and no arginine methyltransferase inhibitor (AdOx) (left) versus AdOx 20 μM (right). Arrows indicate poly-GR and aDMA positive cytoplasmic aggregation (c, d) Effects of AdOx on poly-GR100 cytoplasmic inclusion formation. AdOx did not significantly decrease GFP positive, transfected cell numbers. c AdOx significantly decreased ratio of cytoplasmic inclusions in transfected cells – 43% at 0 mM, 27% at 5 mM and 25% at 20 mM. d Quantitative analysis and representative image showing cytoplasmic inclusions at different concentration of AdOx. Counts were made of over 500 transfected cells for each condition (with at least duplicate counts performed for each field analyzed). All variables analyzed with Kruskal-Wallis ANOVA on Ranks followed by Tukey’s post hoc analysis and data are displayed as median (25th and 75th range). *Statistically significant p-value (p < 0.05); all p-values for ANOVA on Ranks comparison of all three groups. Scale bars in (a) and (b): 20 μm