Fig. 7

SETX rescues delayed recovery of nucleolus fluorescence in cells expressing arginine-containing dipeptide repeats. a We transfected HEK293 cells with a mCherry-nucleolin vector (red), and co-transfected either a BFP-GR(50) expression construct or BFP empty vector, in combination with either a SETX-GFP expression construct or GFP empty vector. After 48 h, we performed fluorescence recovery after photobleaching (FRAP). Here we see representative images depicting the photobleaching and recovery of fluorescence at specific times after the photobleaching the nucleolus of mCherry-nucleolin expressing cells. Scale bar = 2 µM. b Results of FRAP shown at two second intervals from photobleaching (t = 0) until 90 s after photobleaching. Note initial delayed fluorescence recovery in GR(50) expressing cells, and enhanced fluorescence recovery in SETX expressing cells. n = 40 cells/condition. c Quantification of the mobile fraction of the nucleolus from (B). Note the significant reduction in the mobile fraction of the nucleolus in GR(50) expressing cells. ANOVA with post-hoc Tukey test; *P < 0.05. d Measurement of the half-time of fluorescence recovery from (B). Note significantly increased half-time of recovery in GR(50) expressing cells, which is rescued in GR(50) transfected cells co-expressing SETX. ANOVA with post-hoc Tukey test; **P < 0.01. e Analysis of the kinetics of recovery from (B). Note significantly slowed recovery in GR(50) expressing cells, which is rescued in GR(50) transfected cells co-expressing SETX, and also how SETX expression accelerates recovery in control cells. ANOVA with post-hoc Tukey test; **P < 0.01. See Additional file 1: Table S1 for all P values. Error bars = s.e.m