Supplementary Figure 4: SFK(s) mediate the nuclear stiffening to force.

a, Nuclei isolated from Hela cells were incubated with anti-nesprin-1-coated magnetic beads and stimulated with a permanent magnet for different amounts of time. Emerin was immunoprecipitated and its tyrosine phosphorylation was analysed by western blot. All results are representative of at least three independent experiments. b, Nuclei isolated from Hela cells were incubated with anti-nesprin-1-coated magnetic beads and pretreated 30 min with Gleevec (10 μM), SU66056 (2.5 μM) or FAK inhibitor (5 μM). After stimulation with a permanent magnet for 3 min, tyrosine phosphorylation of nuclear proteins was analysed by western blot. All results are representative of at least three independent experiments. c, Nuclei isolated from Hela cells were incubated with anti-nesprin-1-coated magnetic beads and stimulated with a permanent magnet for 3 min. Src expression, Src phosphorylation on Y416, FAK expression and FAK phosphorylation on Y397 were assessed by western blot. All results are representative of at least three independent experiments. d, Change in bead displacement between the first and 6th pulse of force applied to beads coated with anti-nesprin-1 antibody bound to nuclei incubated with no ATP (n = 14 beads) or with SU6656 (n = 22 beads) for 30 min (untreated control n = 13 beads). Displacements were calculated relative to the first pulse of force applied to untreated nuclei (Error bars represent s.e.m., *P < 0.05, data were collected from 3 independent experiments and analysed by two-tailed unpaired t-test).