Cytoplasmic FUS inclusions disrupt the detyrosinated MT network. (A) Percentages of NIH/3T3 cells with Glu-MTs in the presence or absence of cytoplasmic FUS granules. Cells containing any visible Glu-MT fibers were considered Glu-MT–positive. The numbers above each bar indicate the total numbers of cells observed in independent experiments (three for RFP-FUS(R521C), one for RFP-FUS(495X), and four for RFP-FUS(P525L); each against a corresponding RFP control). *, P < 0.0001; Student’s t test. (B) Representative IF images of total α-tubulin or Glu-tubulin in RFP and RFP-FUS(R521C) cells with or without cytoplasmic granules (arrowheads). (C) IF images of total α-tubulin and acetylated-tubulin in GFP and GFP-FUS(P525L) cells with cytoplasmic granules and quantification of relative intensities. The nonsignificant p-value was obtained by Mann–Whitney U test. (D) Representative images of Glu-tubulin staining in disaggregated and aggregated cells after Hsp104 induction, and quantitations of Glu-MT–positive cells from cells expressing the indicated Hsp104 variants. P-values between indicated pairs were P = 0.0599 (A503V) and P = 0.0934 (A503V-DPLF). (C and D) The total number of cells observed in two (C) or three (D) independent experiments is indicated within each bar. Error bars show SEM. Bars, 10 µm. Dox, doxycycline; Wt, wild type.