Live-cell imaging of circular and long noncoding RNAs associated with FUS pathological aggregates by Pepper fluorescent RNA

The lncRNA nHOTAIRM1 colocalizes with physiological and pathological SG. (A) Combination of smFISH and IF for the detection of nHOTAIRM1 (magenta), G3BP1 (gray), and FUS (green) in stressed WT (upper panel) or stressed FUS^P525L (lower panel) iPSCs-derived MNs. Nuclei stained with DAPI are shown in cyan. Scale bars, 10 μm. (B, C) Pie charts representing nHOTAIRM1 percentage of colocalization in WT (B) or FUS^P252L (C) MNs with G3BP1-positive granules (light blue), FUS-positive granules (green), G3BP1- and FUS^P525L-positive granules (gray), and percentage of free nHOTAIRM1 spots (magenta). Mean percentages from three independent biological replicates are shown (N_WT = 549 cells; N_FUS^P525L = 84 cells).