Figure 3.

Immunofluorescent Staining of Nucleoporins in Control and Affected Fibroblasts

Primary fibroblasts derived from affected individual A1 and an unrelated control individual were analyzed by indirect immunofluorescence with focus correction for the mid-plane of the nucleus and images captured as 5×5 tiles and merged into wider fields to allow quantitative analysis. Representative images of staining with several Nup-specific antibodies are shown on the left. Quantitative summaries of the average staining intensity per cell, derived from multiple fields and >100 cells in each category, are shown on the right. NUP214 and NUP88 staining intensity was reduced in the affected individual’s fibroblasts both at the nuclear rim and in the total cellular signal. Bars indicate SEM. See Material and Methods for further details.