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. 2010 Oct 2;285(51):39922–39934. doi: 10.1074/jbc.M110.178087

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Altered interactions of DHC/DIC and dynein/dynactin in Loa. A, antibodies against DHC were cross-linked to cyanogen bromide-activated Sepharose 4B beads to coimmunoprecipitate both the dynein DIC subunits and dynactin from cytosolic extracts of newborn mouse brains. There was no difference between +/+ and Loa/Loa in the precleaned brain homogenates, as control (right panel, Input), but differences between +/+ and Loa/Loa in IP DIC and p150^Glued are present (left panel). B, antibody against dynactin p150^Glued coimmunoprecipitated both DHC and DIC subunits from cytosolic extracts of newborn mouse brains. There was no difference between +/+ and Loa/Loa in the precleaned brain homogenates (right panel), but immunoprecipitated DIC and DHC were different between +/+ and Loa/Loa (left panel). C, ratios of DIC/DHC and p150^Glued/DHC in Loa/Loa were compared with those of +/+ by densitometry quantification of DIC and p150^Glued bands in A. Two-way analysis of variances (ANOVA) and Bonferroni post-tests indicated that there is a significant difference between the genotypes (p < 0.0001, residual degrees of freedom = 19) and that the fold changes in Loa/Loa DIC/DHC and p150^Glued/DHC ratios are significantly different relative to +/+ (p < 0.001 for both ratios). D, DIC/p150^Glued and DHC/p150^Glued ratios in Loa/Loa were compared with those in +/+ by densitometry quantification of DIC, DHC, and p150^Glued bands in B. ANOVA and Bonferroni post-tests showed that the genotype significantly affects the ratios (p < 0.0001, residual degrees of freedom = 22) and that the fold change in Loa/Loa DIC/p150^Glued ratio is significantly different relative to +/+ (p < 0.001). DHC/p150^Glued ratios were not significantly different (p > 0.05), but they showed the same trend as DIC/p150^Glued ratios.