Fig. 6: Kapβ2W460A:W730A prevents and reverses R-DPRs-induced TDP-43 aggregation as efficiently as WT Kapβ2. | Communications Biology

Fig. 6: Kapβ2W460A:W730A prevents and reverses R-DPRs-induced TDP-43 aggregation as efficiently as WT Kapβ2.

From: NLS-binding deficient Kapβ2 reduces neurotoxicity via selective interaction with C9orf72-ALS/FTD dipeptide repeats

Fig. 6

His-SUMO-TDP-43 (5 μM) incubated with buffer, 5 μM WT Kapβ2 or Kapβ2W460A:W730A, in the presence of 10 μM (GR)20 (A), or 10 μM (PR)20 (C). Aggregation was assessed by turbidity measured at 395 nm. Solid lines are normalized mean. Dotted lines of corresponding colors represent ± SEM (n = 3 independent experiments). Quantification of the area under the curve for the curves in (A) and (C) for (GR)20 (B), and (PR)20 (D), respectively. Color code is the same as in (A) and (C). Unpaired Student’s t-tests were used to compare different conditions. E Fluorescence images of His-SUMO-TDP-43 (2 μM) condensates formed in the presence or absence of the indicated R-DPR (2 μM) and the indicated Kapβ2 variant (4 μM). 200 nM His-SUMO-TDP-43-GFP and 100 nM TAMRA-R-DPR were added for visualization. Scale bar: 25 μm. F Indicated Kapβ2 variant (4 μM) was added into His-SUMO-TDP-43 (1 μM) condensates performed in the presence of the indicated R-DPR (1 μM). 2 hours after the addition of Kapβ2, fluorescence images of the samples were taken. 200 nM His-SUMO-TDP-43-GFP and 100 nM TAMRA-R-DPR were added for visualization. Scale bar: 25 μm.

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