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. 2023 Jul 11;42(17):e111719. doi: 10.15252/embj.2022111719

Figure EV5. Nuclear TDP‐43 droplets do not localize to nuclear speckles.

Figure EV5

  1. Representative confocal microscopy images of HEK293 cells stained for the Cajal body marker coilin. Scale bar: 10 μm (5 μm for insets).
  2. Representative confocal microscopy images of HEK293 cells hybridized with a fluorescent NEAT1 probe to mark the paraspeckles. The field overview is shown as a maximum intensity Z‐projection (thickness of ~10 μm, in steps of 0.21 μm). Scale bar: 10 μm (5 μm for insets).
  3. Representative confocal microscopy images of the isogenic HEK293 lines expressing the different GFP‐TDP‐43 variants for 24 h and stained for the nuclear speckle marker pSC35.
  4. 3D analysis quantification showing the percentage of each of the analyzed subnuclear compartments that colocalize with each of the GFP‐TDP‐43 variants as shown in Fig 5E and F; Appendix Fig S5C. N = 8–21 cells. Kruskal–Wallis test with Dunn's multiple comparisons post hoc test.
  5. Representative confocal microscopy images of the isogenic HEK293 lines expressing the different GFP‐TDP‐43 variants for only 4 h to achieve similar expression levels and stained for the Cajal body marker coilin.
  6. 3D analysis quantification showing the percentage of Cajal bodies that colocalize with each of the GFP‐TDP‐43 variants as shown in (E). N = 39–54 cells. Kruskal‐Wallis test with Dunn's multiple comparisons post hoc test. ns: not significant, **P < 0.01, ****P < 0.0001. Violin plots show mean and quartiles.