Extracts were analyzed by WB after a freeze-and-thaw cycle at −20°C (−20°C F/T; A,B) or immediately after preparation (fresh; C,D). Membranes were probed with either anti-Tau phospho S396 (pTau; A–D,F) or anti-Tau [E178] (Total Tau; A–D) antibodies. Extracts were prepared from dorsal hippocampus (A,C) or frontal cortex (B,D) of Wistar (WT) and WAR rats, as discriminated above each lane. (E) pTau/25kDa band ratio. A representative WB comparing extracts from frontal cortex stored at different temperatures is shown (n=3 per group; *P<0.05). Some extracts were additionally probed for β-actin (F). The arrow-head highlights the ∼25 kDa regions on the membranes.