Fig. 2

SETDB1 is required for EpiGo-KRAB-mediated genomic clustering. a Live cell tracking of C19Q dynamics in EpiGo-Control (dCas9) and EpiGo-KRAB (dCas9-KRAB) cell lines. These cell lines were stably expressed PCP-GFP, C19Q-sgRNA-2XPP7, and dCas9 or dCas9-KRAB. The HaloTag was knocked-in at the C-terminus of HP1α and used to examine the colocalization between C19Q (green) and HP1α (magenta). The dCas9 or dCas9-KRAB was induced to express 24 h before tracking. The dynamics of C19Q (green) and HP1α (magenta) were tracked for 240 mins. Images were captured every 30 mins. b 3D-SIM images of C19Q and HP1α in EpiGo-Control and EpiGo-KRAB cell lines. C19Q (green) was labeled by GFP and HP1α (magenta) was labeled by HaloTag. The colocalization between C19Q and HP1α was shown in merged images. c Percentage of C19Q foci clusters in EpiGo-Control (dCas9) and EpiGo-KRAB (dCas9-KRAB) cell lines. C19Q cluster was defined as that all the C19Q loci were confined within 1 μm in each dimension. n = 200 transfected cells in each experiment and the error bar is standard deviation (SD) from three independent experiments. d RT-qPCR analysis of SETDB1 knockdown efficiency. RNA was extracted from the EpiGo-KRAB cells lines transfected with either control siRNA or SETDB1 siRNA. n = 200 transfected cells in each experiment and the error bar is standard deviation (SD) from three independent experiments. e 3D-SIM images of C19Q (green) and HP1α (magenta) in EpiGo-KRAB cell lines in the presence of control siRNA or SETDB1 siRNA. f Percentage of C19Q foci clusters in the U2OS-dCas9-KRAB cells lines transfected with either control siRNA or SETDB1 siRNA. n = 200 transfected cells in each experiment and the error bar is standard deviation (SD) from three independent experiments