Validation of drug treatments and LINC complex disruption. (A) IF validation of actin-cytoskeleton disruption in BJ-5ta cells treated with either cytochalasin B or latrunculin. Phalloidin (green) was used as a marker for F-actin. (B) IF validation of tubulin disruption in BJ-5ta cells treated with nocodazole. (C–F) Western blot validation of siRNA knockdowns of (C) vimentin (D) SUN1 and SUN2 (SUN1/2), (E) SUN1, or (F) SUN2 knockdown in BJ-5ta cells. Tubulin was used as a loading control. (G) Representative fluorescence images of MCF10A cells transiently transfected with ss-GFP-KDEL or ss-SUN1L-GFP-KDEL (green) probed for Nesprin2/4 (red) to assess loss of nesprins from the NE that is indicative of LINC complex perturbation. (H–J) Representative fluorescence images of BJ-5ta cells depleted of (H) Nesprin1, (I) Nesprin2, or (J) Nesprin3 (green), labeled with respective antibodies. Hoescht was used to visualize DNA (blue). Scale bars 10 µm. Source data are available for this figure: SourceData FS2.