Extended Data Fig. 6: YAP regulates TDP-43 phase separation.
From: YAP maintains the dynamics of TDP-43 condensates and antagonizes TDP-43 pathological aggregates
a, MBP–TDP-43 purified from bacteria and mCherry–YAP from DMSO or NaAsO2 treated 293T cells were separated by SDS–PAGE and stained with Coomassie Blue. The experiment was repeated two independently times with similar results. b, Enlarged view of TDP-43 condensates formed in the presence of mCherry or mCherry–YAP. c, HeLa cells transfected with TDP-43-GFP ΔNLS and mCherry–YAP were treated with 250 µM NaAsO2, fixed, and imaged by SIM. Representative images were shown. d, Lysates prepared from HeLa cells treated with λ-phosphatase were subjected to immunoblotting analyses with the indicated antibodies. The experiment was repeated two independently times with similar results. e,f, In vitro phase separation of MBP–TDP-43 (labelled with Alexa 488) in the presence of mCherry–YAP that were purified from DMSO or NaAsO2 treated 293T cells. The reaction was conducted in 50 mM Tris, pH 7.5, 150 mM NaCl and 10% dextran 70. Area occupied by TDP-43 dense phase was quantified (e). Data were shown as mean ± s.d. The experiments were performed for a total of 2 independent times with similar results. 7 images for each condition from one representative experiment were analysed and plotted. g,h, WT HeLa cells expressing TDP-43-GFP ΔNLS were transfected with siCtrl or TEAD1/3/4 and subjected to FRAP. The fluorescence intensity was monitored over time and quantified (h). The experiments were performed for a total of 2 independent times with similar results. 15 condensates from one representative experiment were analysed and plotted. Data were shown as mean ± s.d.
