Immunoblots of sarkosyl-insoluble tau from affected members from HFTD1. (A) Immunoblots of sarkosyl-insoluble tau extracted from hippocampus from an Alzheimer’s disease patient (lane 1) and hippocampus (lanes 2, 3) and frontal cortex (lanes 4–6) from three affected members of HFTD1 stained by antiserum BR134. Tau bands of 60, 64, 68, and 72 kd are indicated. (B) Immunoblots of sarkosyl-insoluble tau from brain of an affected HFTD1 member, before (lane 1) and after (lane 2) alkaline phosphatase treatment, using antiserum BR133. Note that before alkaline phosphatase treatment two major bands of 64 and 68 kd and a minor band of 72 kd are present; after alkaline phosphatase treatment, three major bands are visible which correspond to tau isoforms with 4 repeats and no amino-terminal amino acid insert (4R) and both 4 and 3 repeat tau isoforms with 29-amino acid amino-terminal insert (4R + 29aa and 3R + 29aa). These are isoforms D, E and B respectively in panel (C). (C) Schematic representation of the 6 tau isoforms present in adult human brain showing the position of G272V and P301L mutations. The P301L mutation is in the alternatively spliced exon 10 and is present only in tau isoforms with 4 repeats, whereas G272V is present in all isoforms.