Fig. 6

VEX-complex interactions impact abundance and location. a, b Protein-blot and immunofluorescence analysis following Tet-induced VEX1 knockdown and Tet-induced ^mycVEX1 overexpression, respectively (48 h). In the upper immunofluorescence panels, cells were stained with α-VSG-2 and α-VSG-6 to confirm the dual VSG expression phenotypes reported previously¹⁷. In the lower panels, cells were stained with α-myc for VEX2 localisation (a) or both α-myc and α-GFP for VEX1 and VEX2 localisation, respectively (b). The bar graphs indicate the proportion of ≥300 nuclei with 1–2 ^mycVEX2 foci following 48 h of VEX1 knockdown (a) or with 1–2 ^GFPVEX2 foci following 48 h of VEX1 overexpression (b). The proportion of nuclei displaying a distributed VEX1 signal (due to overexpression) is also shown in b; the remaining cells displayed no detectable signal. c Protein-blot and immunofluorescence analysis of VEX1^Myc following Tet-induced VEX2 knockdown (24 h). The bar graph indicates the proportion of nuclei with 1–2 VEX1^myc foci or distributed signal, the remaining cells displayed no detectable signal. The bean-plot shows signal intensity in ≥100 nuclei with 1–2 major VEX1^myc foci (−Tet) or nuclei with a distributed signal (+Tet 24 h) following VEX2 knockdown. Black lines show the medians; pink/red lines represent individual data points; polygons represent the estimated density of the data. CTNF corrected total nuclear fluorescence, normalised to the nuclear area. EF1α was used as a loading-control. DNA was counter-stained with DAPI; scale bars 2 μm. The images in (c) correspond to maximal 3D projections of 0.1 μm stacks. For bar graphs, data representative of independent experiments and averages from two independent biological replicates; error bars, SD. ns, non-significant; *p < 0.05; ***p < 0.001; ****p < 0.0001 (two-tailed paired and unpaired Student’s t tests applied to bar graphs and bean plot, respectively). Source data are provided as a Source Data file