Fig. 3

Hmga2 suppression causes a remarkable distortion of the nuclear lamina. A Lmnb1 immunofluorescence (red) on Hmga2 wt and KO cells at day 1 of EpiLC induction, showing the distortion of the NL in Hmga2 KO cells. Maximum projection of z-slices (ROI 1024×1024) is shown; quantification graphs obtained by counting >200 nuclei/condition. B Representative images (single Z-plane, ROI 1024×1024) of the nuclear abnormalities (nuclear protrusions and nuclear blebbing) observed in Hmga2 KO cells at day 1 of EpiLC induction and absent in wt cells; quantification graphs obtained by counting >200 nuclei/condition. C Three-dimensional analysis of z-slices maximum projections showing the distortion of Lmnb1 (red) as well as the reduced porosity (grey dots inside the NL) detected in Hmga2 KO cells compared to the wild-type ones. Scale bar= 10 μm. D Lmnb1 immunofluorescence (red) on Hmga2 KD ESCs induced into EpiLCs (day 3) showing an evident distortion of the NL, with some enlarged and wrinkly nuclei. Non-silencing siRNA was used as control. Quantification graphs obtained by counting >400 nuclei/condition. E Lmnb1 immunofluorescence on Hmga2 KD cells showing the nuclear abnormalities (nuclear blebbing, enlarged nuclei) typical of cells devoid of Hmga2. DAPI (blue) was used to counterstain the nuclei. A single plane of the z-stack projection is shown. Scale bars of A, B, D, and E = 50 μm. Error bars represent standard deviation. Statistical significance on three biological replicate experiments was determined using Student’s t-test (ns: not significant, *p<0.05, **p<0.01)