Fig. 6
From: Activated cofilin exacerbates tau pathology by impairing tau-mediated microtubule dynamics
Active cofilin interacts with tubulin and disrupts microtubule dynamics. a, b Hela-V5-Tau cells were transfected with mRFP, cofilin-mRFP, cofilin-S3A-mRFP, or cofilin-S3E-mRFP for 48 h, treated with nocodazole for 30 min and recovered for another 30 min (scale bar = 10 μm). White boxes magnified to the right. b Quantification of transfected cells with microtubule organizing center (MTOC) normalized to mRFP controls. Data are expressed as mean ± SEM (one-way ANOVA with Tukey post hoc, n = 4 repeats, #p < 0.0001). c, d Hela-V5-Tau cells were transiently transfected with control, cofilin-mRFP, cofilin-S3A-mRFP, or cofilin-S3E-mRFP mutants. Cells were lysed with 1% CHAPS buffer, and lysates were immunoprecipitated for tubulin and immunoblotted for cofilin. Representative blot shows cofilin-S3A preferentially interacts with tubulin compared to cofilin-S3E. d Quantification of tubulin–cofilin complexes. Data are expressed as mean ± SEM (one-way ANOVA with Tukey post hoc, n = 4, **p = 0.0016, ***p = 0.0003). e, f Hippocampal primary neurons derived from WT and Tau-P301S were transduced with mRFP, cofilin-mRFP, cofilin-S3A-mRFP, or cofilin-S3E-mRFP adenovirus on DIV7 and subjected to immunocytochemistry for detyrosinated tubulin on DIV21 (scale bar = 20 μm). f Quantification of detyrosinated tubulin intensity. Data are expressed as mean ± SEM (one-way ANOVA with Tukey post hoc, n = 15–20/genotype **p = 0.001, *p = 0.0228 compared to WT RFP control)
