Fig. 6

Changes in chromosome structures and localization patterns of condensin after knockdown of cohesin components. a,g Fluorescence images of metaphase chromosomes. The chromosomes were immunostained with antibody against SMC4 and DAPI (left) in ESCs and MEFs. Scale bar = 2.5 μm. b,h 3D SIM images of metaphase chromosomes. The cells were stained with anti-SMC4 antibody and PI. Scale bar = 2.5 μm. c,i 3D-rendering images of SMC4-stained metaphase chromosomes. SMC4 is pseudo-colored in each color to measure the localization and intensity of condensin on metaphase chromosomes. 3D-rendered images were created using the Nikon NIS software. Scale bar = 2.5 μm. d Intensity and distribution of SMC4 in metaphase chromosomes. e,f,j,k The intensity and spatial density of SMC4 per chromosome area in metaphase ESCs (e,f) and MEFs (j,k). The y-axis values represent the intensity of SMC4 and the spatial density of SMC4 per a chromosome area. The y-axis values were calculated by dividing the volume of chromosome area by the intensity or volume of SMC4 spots. The area and intensity were measured using Nikon NIS software. Each biological replicate is color-coded (red, green, and blue) and the average of each replicated data is indicated with a larger dot, and black bars indicate the averages of three means (n = 30 for condition). The error bars are the mean ± SD from three independent biological replicates. Statistics: Paired two-tailed t-test, ns: not significant, *P < 0.5, **P < 0.01, and ***P < 0.001