Fig. 5.

STEF regulates nuclear re-orientation and height. a Spinning disc confocal images of either control MEFs (Control), or MEFs depleted for endogenous STEF (STEF KO) or depleted for endogenous STEF, but re-expressing exogenous wild-type STEF (STEF KO + WT STEF) plated on collagen-coated cross-bow micropatterns, fixed after 7 h and stained for nuclei (DRAQ5) and actin (Phalloidin). b Schematic of the front-rear polarised morphology and nuclear position on micropatterns also showing how nuclear angles were measured. c Histogram showing quantification of nuclear angles relative to x axis in MEFs of a. Representative replicate from two independent experiments. Analysis conducted using the 'fit ellipse' tool on ImageJ. d Representative immunofluorescence images of MEFs plated as in a. Right panels, maximal projection images of the x–z plane of the nucleus. e Quantification of nuclear height of MEFs of d. Image z-stacks for DRAQ5 stain were imported into the Imaris software and the shortest principal axis of the nucleus (height) was measured. Values represent the mean of three independent experiments, >100 cells per condition, per replicate. Statistical significance was verified using a one-way ANOVA, using Dunnett’s multiple comparison test to compare the means of each sample to the control. * p < 0.05, n.s. = not significant. Error bars represent S.E.M. Scale bars = 10 µm throughout