Figure 9.

β2-spectrin organization on fiber cell membranes is disrupted in the absence of Tmod1, but β2-spectrin levels and membrane association are unchanged. (A) Single optical sections or (B and C) 3D images of Z stacks of β2-spectrin and F-actin showing matched regions of fiber cells from 1-mo-old mouse lenses as in Fig. 8. En face views of vertices and narrow sides (B) or broad sides (C). Bars: (A and B) 4 µm; (C) 2.2 µm. (D) Western blots of β2-spectrin, N-cadherin, actin, Tmod1, and Aquaporin 0 (AQP0) in total cortical fiber cell extracts. Each lane is 1/4 of a 1-mo-old lens cortex from a different animal. (E) Quantification from blots in D, normalized to AQP0. Levels of β2-spectrin, N-cadherin, and actin are not different in absence of Tmod1 (P < 0.24, 0.36, and 0.2, respectively). n = 4 animals per genotype. (F) Western blots of β2-spectrin, N-cadherin, Tmod1, actin, and γTM in cortical fiber cell total extracts (T), cytosol (C), membranes (M), Triton X-100 supernatant (TxS), and pellet (TxP) from 8-mo-old mice.