Fig. 2

SUN1 and lamin B1, but not lamin A, contribute to TGFβ1-induced nuclear deformation. A Immunoblotting results of the indicated proteins in Huh7 cells transfected using the indicated siRNAs for 24 h then treated with 10 ng/mL TGFβ1 for 3 days. C, control; LA, lamin A; LB1, lamin B1; S1, SUN1; S2, SUN2; Eme, Emerin. ACTIN was used as a loading control. B Confocal images of cells transfected with the indicated siRNAs for 24 h, followed by TGFβ1 treatment for 48 h. Cells were immunofluorescent stained with lamin B (white) and the nuclei were counter stained with Hoechst 33342 (blue). C Categorization of the nuclear shape in cells treated using the methods in (B). Number of cells quantified under each experiment condition was denoted. *P < 0.0001, Fisher’s exact test. D Confocal images of cells transfected with siRNAs against SUN1 or lamin B1 for 24 h, followed by TGFβ1 treatment for 48 h. Cells were immunofluorescent stained with rabbit anti-SUN1 and goat anti-lamin B. The nuclei were counter stained with Hoechst 33342. E Confocal images of cells with mock or TGFβ1 treatment for 48 h, followed by 1 μM treatment of cytochalasin D and/or nocodazole for 30 min. Cells were immunofluorescent stained with phalloidin (to denote F-Actin), mouse anti-α-tubulin and goat anti-lamin B. Nuclei were counterstained with Hoechst 33342. Act, F-actin; tub, α-tubulin, LMNB, lamin B. All images are the sum of z-stacks