Figure 4.
The G1073S variant decreases the actin-binding affinity of SHROOM3 protein. (A) FLAG-tagged BN, FHH, or ∆G1073S SHROOM3 proteins were overexpressed in HEK293 cells, followed by immunoprecipitation against FLAG. Immunoprecipitated lysates were immunoblotted using antibodies against FLAG, ROCK1, and ACTB. A representative Western blot of immunoprecipitated lysate is provided. (B) Quantification of the Western blot showed that FHH SHROOM3 and ∆G1073S mutant had significantly reduced actin-binding affinity compared to BN SHROOM3. (n = 3 per group. [*]P < 0.05 vs. BN.) (C) ROCK1-binding affinity was not different among the three alleles (n = 3 per group).
