Fig. 6
From: NDP52 activates nuclear myosin VI to enhance RNA polymerase II transcription
NDP52 transcription regulation. a Immunofluorescence staining against NDP52 (green), RNAPII (magenta) and DNA (cyan), as performed in Fig. 1b. Insert depicts colocalising foci. Scale bar 10 μm for whole images and 1 μm for inserts. b Immunofluorescence on HeLa nuclei as in (a) with Pearson’s coefficient (error bars represent SEM from 10 images). c Co-immunoprecipitation of NDP52 and RNAPII, using an antibody against NDP52 to immunoprecipitate RNAPII from HeLa cells. Immunoprecipitation does not occur in HeLa cells subjected to siRNA knockdown of NDP52. d Western blot against RNAPII following isolation from nuclear extracts by NDP52 constructs and CoCoA. Loading controls for the recombinant proteins is shown in Supplementary Fig. 9. e Pull-down of CoCoA by His-tagged NMVITAIL both at 5 μM. P and S represent pellet and supernatant fractions. f In vitro transcription by HelaScribe extracts following antibody depletion as described in Methods. Samples were normalised to a non-depleted control reaction (error bars represent SEM **p < 0.001 by two-tailed t-test) from 5 independent experiments (Supplementary Fig. 8a, b)
