Fig 6. RAC1 interacts with FLNA in the cytoplasm and deletion of RAC1 reduces the production of cleaved C-terminal fragment of filamin A (FLNA^CT).

(A) Expression of RAC1 (green) and FLNA (red) is co-localized mainly in the cytoplasm of BMMs (yellow) as detected by immunofluorescence staining. Scale bar represents 10 μm. (B) Co-immunoprecipitation identifying FLNA^CT as an interacting partner of RAC1. Total proteins obtained from BMMs immunoprecipitated with FLNA^CT and then immunoblotted with RAC1 antibodies (upper) or vice versa (lower). IgG served as negative controls. Full-length of FLNA and FLNA^CT were indicated and actin served as internal loading control. (C) Immunoblotting of FLNA in Rac1^fl/fl and Rac1^fl/fl/LC BMMs. (D) Immunofluorescence staining of FLNA^CT in Rac1^fl/fl and Rac1^fl/fl/LC BMMs. Quantification of nuclear FLNA^CT expression in Rac1^fl/fl or Rac1^fl/fl/LC BMMs. Scale bar represents 20 μm. Mean ± SEM values of at least quadruplicated experiments. Student’s t-test was used. *p<0.05, **p<0.01.