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Fig. 7. Nesprin-2 localisation is affected in cells where Sun1 expression has been silenced by siRNA. (A-C) HeLa cells expressing stably V5-tagged hSun1 and HaCaT cells (D-L) were transiently transfected with a combination of plasmids (pJG173/174) encoding siRNAs targeting hSun1. The distribution of Sun1 (panel A, anti-V5; panels D,G,J, anti-Sun1 281 serum), lamin A/C (panels B and E) and Nesprin-2 (panel H, mAb K20-478; panel K, mAb K49-260) was investigated by indirect immunofluorescence in knockdown cells (indicated by asterisks). In Sun1 knockdown cells, the lamin A/C localisation remained unaltered (B and E), whereas Nesprin-2 staining was either absent or reduced. DNA was stained by DAPI. The images shown were taken by confocal microscopy and merged to visualise colocalisation (panels C,F,I,L). Bars, 10 µm.





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