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. 2015 Apr 10;10(4):e0123018. doi: 10.1371/journal.pone.0123018

Fig 6. Role of FLNA in, and effect of FLNAC on, the interaction of PC2 with actin.

Fig 6

A, effect of FLNA KD on the PC2-actin interaction revealed by co-IP with anti-GFP (EU4) antibody in A7 PC2 stable cells. B, effect of FLNA KD on the PC2-actin interaction revealed by co-IP with PC2 (H-280) antibody in HeLa cells. C, localization of PC2, filamin-A and actin in A7 (upper panel) and IMCD (lower panel) cells, and co-localization of PC2 and Na/K ATPase in A7 cells were determined by immunofluorescence assays, as previously described [21]. GFP-PC2 are stably expressed in A7 and IMCD cell lines. Primary antibodies against FLNA (H-300), β-actin (C-4), and Na/K ATPase (H-300) were used. Rabbit Cy3- and mouse Cy5-conjugated secondary antibodies were used to detect FLNA (or Na/K ATPase) and actin, respectively. Images were acquired using AIVI spinning disc confocal microscopy with x60 objective. D, effect of FLNAC on the FLNA-mediated PC2-actin interaction using A7 and M2 (as control, no FLNA) cells revealed by biotinylation assays that showed recruitment of actin to the PM.