Tmod3 levels in Tmod1-null ghosts are one-fifth that of Tmod1 in ghosts from wild-type mice, but levels of actin, TM, and other actin-binding proteins are unchanged. (A) Western blots of Tmod1, Tmod3, band 3, 4.1R, dematin, actin, TM5NM1, and EcapZβ2 in Mg⁺⁺ ghosts from wild-type and Tmod1-null mice. Each lane is from a different individual animal. (B) Ratios of proteins to band 3 determined by densitometry from Western blots in panel A, showing no significant differences between wild-type and Tmod1-null samples (4.1R, P < .8; dematin, actin, TM5NM1, and EcapZβ2, P < .2). (C) Western blots of α- and β-adducin in Mg⁺⁺ ghosts from 2 wild-type and 2 Tmod1-null mice, using Coomassie blue staining for spectrin from a duplicate gel as a loading control. Adducin levels are unaffected by the absence of Tmod1. (D) Molar ratio of actin/Tmod1 in wild-type ghosts and actin/Tmod3 in Tmod1-null ghosts. Nanograms of actin and Tmod1 or Tmod3 were quantified per microliter of ghosts by Western blotting according to standard curves for each purified protein, as shown in supplemental Figure 1.