Extended Data Fig. 2: SETD2 loss during G2/M leads to nuclear lamina defects.
From: A SETD2–CDK1–lamin axis maintains nuclear morphology and genome stability
a, Representative confocal microscopy images of control and SETD2-KD HKC cells immunostained with anti-lamin A/C and lamin B1 antibodies. DNA counterstained with DAPI. b, Representative confocal microscopy images of control and Setd2-KO MEF cells immunostained with anti-lamin B1 antibody. DNA counterstained with DAPI. c, Schematic of cell synchronization by double thymidine block and release method used in d–f. d, Immunoblot analysis of indicated proteins in untreated and dTAG-47 treated (at 6 h post release) cells. e, Representative images of either untreated or dTAG-47 treated SETD2–FKBP cells, fixed at the indicated timepoints post release from thymidine block and immunostained with anti-lamin A/C antibody. DNA counterstained with DAPI. f, Quantification of nuclear defects observed in e. Ordinary one-way ANOVA with Sidak’s multiple comparison test was performed on data from two independent biological replicates, with n (left to right) = 614, 558, 735, 654, 754, 693 cells; mean ± s.e.m. g, Schematic of cell synchronization by serum starvation of Setd2 (w/flox) MEFs. h, Immunoblot analysis of indicated proteins in untreated and tamoxifen treated (Setd2-KO) MEF cells upon release from G0/G1 block. i, Representative images of either control or tamoxifen-treated MEF cells at the indicated timepoints post release from serum starvation, immunostained with anti-lamin A/C antibody. j, Quantification of nuclear defects from data in i. Two-way ANOVA with Sidak’s multiple comparison test was performed on data from three independent replicates with n (left to right) = 184, 166, 247, 223, 256, 247, 387, 348 cells; mean ± s.d. All scale bars are 10 µm.
