Fig. 3
From: NDP52 activates nuclear myosin VI to enhance RNA polymerase II transcription
Regulation of myosin VI backfolding. a FRET titration of NMVITAIL against CBD (1 μM) or CBD(W1221A) in the presence of DNA, or NDP52, at 10 μM. Data fitting was performed as described in Methods. NDP52 and CBD(W1221A) data are plotted as disconnected points due to low signal change. See Supplementary Fig. 4a–c for raw intensity data. b Representative fluorescence spectra of 2 μM GFP-MVITAIL-RFP ± 15 μM NDP52. c FLIM measurements of in vivo MVI conformational dynamics. Above, schematic showing the constructs. Left, GFP intensity and lifetime images. Right, intensity and lifetime images of the biosensor. Lifetime (τ) is represented by a pseudocolor scale. Scale bar is 20 μm. d Pull-down of CBD by His-tagged NMVITAIL both at 10 μM ± 20 μM 40 bp DNA. P and S represent pellet and supernatant fractions. e Fluorescence anisotropy titrations of MVI domains against a 40 bp FAM DNA, as performed in Fig. 2a
