Comparison of cytosol/membrane levels for Tmods and other actin-binding proteins in wild-type or Tmod1-null RBCs. (A) Coomassie blue–stained sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE) of total RBC extracts (T), RBC cytosol (C), and washed membranes (M) prepared by hypotonic lysis of RBCs isolated from wild-type and Tmod1-null mice. (B) Western blots of Tmods, actin, TMs, dematin, 4.1R, and band 3 in total RBC extracts, cytosol, and washed membranes. Tmod1 and Tmod3 are predominantly associated with membranes, and associations of actin, TMs, dematin, and 4.1R with membranes are not altered in the absence of Tmod1. A small amount of the upper TM5NM1 but not the lower TM5b band is detected in RBC cytosol from Tmod1-null mice.