Supplementary Figure 5: Characterization of lamin A/C acetylation in MEFs.

a) Left; immunoblotting on subcellular fractions of WT/K417R OST-lamin A MEFs. Right; Nuclear abnormalities quantification; Two-tailed unpaired t-test on n=3 independent biological replicates. b) Relative mRNA expression (against Rplp0) of the OST-tagged WT, K97R, K108R, K311R and K486R derivatives (n=3 independent biological replicates) of Supplementary Fig. 5c. c) Nuclear abnormalities quantification in MEFs expressing the indicated lamin A derivatives. Statistical comparisons against WT lamin A using ordinary one-way ANOVA on n=3 independent biological replicates. d) Relative mRNA expression (against Rplp0) of the OST-tagged WT, K311R and K311Q lamin A derivatives (n=3 independent biological replicates) of Fig. 3c. e) Immunoblotting on the indicated subcellular fractions of MEFs expressing WT, K311R or K311Q lamin A. f) Left; Immunoblotting on whole cell extracts of immortalized Lmna KO MEFs expressing WT, K311R or K311Q OST-lamin A. Right; Nuclear abnormalities quantification. Two-tailed unpaired t-test on n=3 independent biological replicates. g) Representative immunostaining for OST-tag (green) and lamin B1 (red) on control and Mof KO K319R/Q MEFs. h) Nuclear abnormalities quantification of Supplementary Fig. 5g. Ordinary one-way ANOVA on n=3 independent biological replicates. i) Representative immunostaining for WT/K59R GFP-lamin B2 (green) and lamin B2 (red). j) Nuclear abnormalities quantification in WT/K59R lamin B2 MEFs. Two-tailed unpaired t-test on n=3 independent biological replicates. k) TEM analysis of paracrystalline arrays. K311R and WT lamin C show similar striped patterns and structural organization of the paracrystalline arrays. Scale bar, 20 nm. l) Quantification of total RNA Pol II and RNA Pol II-S2P. Nonparametric one-way ANOVA followed by a Kruskal-Wallis test. n numbers are stated in the graph and represent nuclear structures counted. m) left:Representative immunostaining of NUP153 in WT and K311R OST-lamin A MEFs. Right: Quantification of NUP153 in main nuclei and nuclear abnormalities of K311R MEFs. Two-tailed nonparametric Mann-Whitney test. n numbers are stated in the graph and represent nuclear structures counted. n) H4K16ac intensities on MEFs expressing WT, K311Ror K311Q OST-lamin A. Means were compared by a Kruskal-Wallis test on n=3 independent biological replicates. The lower and upper hinges correspond to the first and third quartiles. The whiskers extends from the hinge to the largest or smallest values respectively, no further than 1.5 * IQR (inter-quartile range) from the hinge. The middle line represents the median (the middle value of the dataset). Data beyond the end of the whiskers are plotted individually. Data shown are the mean ± s.e.m for a, b, c, d, f, h, j, l, m. For g, i, m dsDNA was counterstained using DAPI and scale bars represent 5μm. Uncropped blots of a, e, f can be found in Supplementary Fig. 9.