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. 2014 Dec 24;290(7):4059–4074. doi: 10.1074/jbc.M114.627919

FIGURE 1.

FIGURE 1.

In situ immunodetection of Tau and characterization of novel monoclonal antibodies in yeast. A, in situ immunodetection of total Tau (TAU5), the pathogenic Tau conformation (MC1), and oligomeric Tau (TOC1) in spheroplasts of the BY4741 wild-type strain expressing human Tau-2N/4R. Scale bar, 2 μm. B, Western blot analysis of protein extracts made from the BY4741 wild-type strain with the empty vector (lane 1), the BY4741 wild type strain expressing Tau-2N/4R (lane 2), the Δmds1 strain expressing 2N/4R Tau (lane 3), and the Δpho85 strain expressing human Tau-2N/4R (lane 4). TAU5 was used as a control antibody. Exposure times were 10 s for ADx215, 2 min for ADx201 and ADx210, and 1 min for TAU5. The open arrows indicate oligomeric Tau as detected by ADx210 and ADx215. The inset for ADx210 shows that this mAb also recognizes monomeric Tau at longer exposure times. C, Western blot analysis of AP-treated samples. Lane 1, extracts of the Δpho85 strain transformed with the empty vector. Lane 2, untreated extracts of the Δpho85 strain expressing Tau; lanes 3–6, the same extract but treated with SAP for 30, 60, 90, and 120 min, respectively.