Validation of LINC perturbation on NCT using the LEXY reporter. (A) Representative fluorescence images of MEFs transfected with either GFP-KASH4 or GFP-KASH4 mutant that cannot bind the SUN domain (yellow arrows) labeled with antibodies against Nesprin3 to validate loss of nesprin from the NE in the presence of dominant-negative GFP-KASH4 but not the mutant control. Scale bar 10 µm. (B) Quantification of LEXY nuclear localization during photoactivation in MEFs transiently transfected with either GFP-KASH4 or GFP-KASH4 mutant to disrupt the LINC complex (GFP-KASH4 n = 14, GFP-KASH4 mutant n = 11). (C and D) Import and export rates from A.