Fig. 9
VEX-complex reassembly and inheritance requires CAF-1. VEX1myc (a), but not mycVEX2 (b), is distributed following tetracycline (Tet) induced CAF-1b knockdown (12 h), as assessed by immunofluorescence analysis. DNA was counter-stained with DAPI; scale bars, 2 μm. The images correspond to maximal 3D projections of 0.1 μm stacks. Error bars, SD; ns, not significant; **p < 0.01 (two-tailed paired Student’s t test). Values are averages from three independent biological replicates and are representative of independent experiments (the remaining cells displayed no detectable signal). c Protein-blot analysis of CAF-1bmyc, mycVEX2 and VEX1myc expression following CAF-1b knockdown. EF1α was used as a loading-control. d–i RNA-seq analysis following CAF-1b knockdown (24 h). d Total number of reads across CAF-1b locus in the parental population or following CAF-1b RNAi. e–i Values are averages from two independent RNAi strains relative to wild-type controls. Panel e shows transcript abundance at the silent VSG-6 ES. Red circles, promoters; red boxes, VSGs; grey boxes, other genes. RPKM/RPM, reads per (kilobase per) million (e–i). Panels f–i highlight different gene cohorts and numbers in square brackets indicate increase/decrease in total transcript abundance for each category; red square, VSG-2; black circle, CAF-1b. Promoter-proximal VSGs and ESAGs (6, 7 and 10) are within 5 kbp of a promoter, distal VSGs are 46 +/−4 kbp from the promoter. Source data are provided as a Source Data file
