Figure 3 | Scientific Reports

Figure 3

From: Mutations in bassoon in individuals with familial and sporadic progressive supranuclear palsy-like syndrome

Figure 3

Western blot analysis of tau and wild-type BSN (BSN[Wt]) or mutated BSN (BSN[Mut]). (A) Protein assay of tau by overexpression of BSN(Wt) or BSN(Mut). HEK293T cells overexpressing cMyc-tagged tau, cMyc-tagged tau and EGFP-tagged BSN(Wt), and cMyc-tagged tau and EGFP-tagged BSN(Mut) were used. Western blot analysis of tau with cMyc-tagged tau and EGFP-tagged BSN(Wt), compared with HEK293T cells overexpressing cMyc-tagged tau, revealed the reduced accumulation of tau bands, especially in the insoluble fraction. However, in Western blot analysis of tau with cMyc-tagged tau and EGFP-tagged BSN(Mut), compared with HEK293T cells overexpressing cMyc-tagged tau, there was no reduction of tau bands in neither the insoluble nor soluble fraction. In this study, tau protein with 4 repeats was used. (B) Comparison of insoluble tau and soluble tau using ImageJ analysis showed that HEK293T cells overexpressing EGFP-tagged BSN(Mut) accumulated more insoluble tau than HEK293T cells overexpressing EGFP-tagged BSN(Wt). Data are means ± S.D. of values from three independent experiments. (C) HEK293T cells overexpressing EGFP-tagged BSN(Wt) and EGFP-tagged BSN(Mut) were used. Western blot analysis of BSN(Mut) compared with BSN(Wt) showed the accumulation of BSN in the insoluble fraction. GAPDH was used as an internal control. (D) Comparison of insoluble BSN and soluble BSN using ImageJ analysis showed the accumulation of BSN in the insoluble fraction and reduced degradation in HEK293T cells overexpressing EGFP-tagged BSN(Mut). Data are means ± S.D. of values from three independent experiments.

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