Fig. 5.

TMEM106B knockdown leads to increased propensity for TDP-43 aggregate formation in an inducible cell system that expresses mislocalized TDP-43 protein. a-d Representative images of double-label immunofluorescence analysis to detect insoluble TDP-43 proteins in iGFP-NLSm cells in the absence (a, b) or presence (c, d) of doxycycline after 72 hours of TMEM106B knockdown. Triton X-100 was used to extract soluble proteins prior to immunolabeling. Green = GFP, reflecting exogenously-expressed GFP-tagged NLSm TDP-43; red = TDP-43 detected with the C1038 anti-TDP-43 antibody. RIPA-insoluble TDP-43 aggregates were detected after doxycycline induction of iGFP-NLSm cells, with TMEM106B knockdown leading to increased numbers of cells with insoluble TDP-43 aggregates. Scale bar = 50 μm. e Quantification of cells with TDP-43 aggregates at 48 and 72 hours after TMEM106B knockdown (n = 3 independent experiments). * One-way ANOVA (TMEM106B knockdown vs. control), with Tukey posthoc pairwise comparison p < 0.05 corrected for multiple comparisons.