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All mice were allowed to sit on the platform for 10 sec after each training trial. Escape latencies, distance traveled, swim paths, swim speeds, and platform crossings were recorded automatically for subsequent analysis. The escape platform was placed in the middle of the channel. If a mouse was not able to find and mount the platform during 60 sec trial, it was guided to it and allowed to sit for up to 10 sec. On the test day, animals were placed in the rectangular test chamber and allowed to explore two identical objects (7.6 cm high x 5.1 cm wide) for a 15 minute training period. For probe trials, the platform was removed and each mouse was allowed to swim for 60 sec. The platform location remained the same throughout training whereas the drop location varied randomly between the four daily trials as well as across the 4 days of training. For these experiments, animals were handled for 3 days and habituated to the chamber (48 cm.times.38 cm.times.20 cm) on the day prior to the test session.

Mice received 2 sessions per day for 4 consecutive days. Primary hTau neuronal cultures were cultured with LNA oligomers until 13 days post plating (DIV 13). On DIV 13, the cultures were rinsed with Dulbecco’s phosphate-buffered saline lacking calcium and magnesium (DPBS, Invitrogen) and fixed in 4% paraformaldehyde/4% sucrose/DPBS for 15 min. One day post plating (DIV 1), half of the supplemented neurobasal (NB) media on the primary hTau mouse embryonic forebrain neuronal cultures was removed and replaced with supplemented NB media containing various concentrations of LNA oligomers. Each of the oligomers targeting the 3′ UTR of an MAPT transcript was tested for its ability to decrease Tau protein in mouse primary neurons expressing the entire human MAPT gene as a bacmid containing transgene (C57-b16 BAC-Tg hTau; Polydoro et. Primary neurons were generated by papain digestion according to manufacturer’s protocol (Worthington Biochemical Corporation, LK0031050). Potency values were generated using wells treated with a broad concentration range of LNA oligomer, from which the resulting normalized Tau/Tuj-1 and Tuj-1 values were analyzed compared to normalized values from saline control samples.

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