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. 2010 Nov 15;286(3):2194–2204. doi: 10.1074/jbc.M110.186924

FIGURE 8.

FIGURE 8.

Effect of point mutations in the LRR domain on Tmod1 pointed end localization. Neonatal rat cardiomyocytes expressing GFP-Tmod1 mutants V232D/F263D/L313D, V232D, F263D, L313D, V232D/F263D, and V232D/L313D were stained for GFP (a, e, i, m, q, and u), F-actin (b, f, j, n, r, and v), and α-actinin (c, g, k, o, s, and w). d, h, l, p, t, and x show merged images of the triple staining (GFP, green; F-actin, red; α-actinin, blue). No localization was observed for GFP-Tmod1 triple mutant V232D/F263D/L313D (a). Consistent localization of GFP-Tmod1(V232D) was only observed in 52.0 ± 5.3% of cells (e, arrowheads), whereas 70% of cells expressing GFP-Tmod1(F263D) and GFP-Tmod1(L313D) displayed consistent pointed end localization (71.3 ± 3.1% and 68.7 ± 2.3%, respectively) (i and m). Cells displaying consistent localization were strikingly decreased when the double mutation was expressed (13.3 ± 7.6% and 10.7 ± 7.0, respectively) (q and u). Nuclear localization (N) was observed in the cells expressing GFP-Tmod1 mutants V232D/F263D/L313D, V232D/L313D, and L313D, suggesting that leucine 313 is responsible for cytoplasmic localization of Tmod1. Scale bar = 10 μm.