Additional file 1: of Partial Tmem106b reduction does not correct abnormalities due to progranulin haploinsufficiency

Figure S1. Measurement of Glycosylated HexA and GCase by Western Blot. The western blots in fig. 3 measured the glycosylated forms of HexA and GCase, which was confirmed by loss of these bands after digestion with the glycosidase PNGase F. The glycosylated forms of HexA and GCase are labeled with black arrows, the unglycosylated forms with gray arrows, and nonspecific bands with asterisks. (DOCX 195 kb

Brain tryptophan.

<p>Brain tryptophan content in µg/mg tissue (a) prefrontal cortex (b) frontal cortex and (c) hippocampus. BALBc and C57 mice received two treatments by gavage of TRP+, TRP− mixtures or water vehicle at 30 minute intervals followed by saline or NSD 1015 to inhibit amion acid decarboxylase at 2 hours. Animals were killed 2.5 hours after the first treatment. Data are collapsed for NSD treatment as no significant effects were predicted or observed. N = 11–12/group. * different from Water. +different from TRP+, ♦ different from corresponding group in BALBc mice. TRP+ and TRP− effectively raised and lowered TRP respectively, although the depletion condition was more consistently effective than the control condition. Similar effects were detected in both strains.</p

Time Course of Behavioral Experiment.

<p>Time Course of Behavioral Experiment.</p

Time course of ATD Moja-De in both strains.

<p>(a) Plasma = plasma tryptophan (µg/ml plasma) (b) HPC TRP = tryptophan in hippocampus (µg/mg tissue) (c) HPC-5-HIAA = 5-HIAA in hippocampus (ng/mg tissue). N = 3–4/group for C57, N = 4–8/group for BALBc. * different from Time 0 and ♦ different from corresponding group in BALBc mice.</p

Brain 5-HT.

<p>Brain 5-HT content in ng/mg tissue in (a) prefrontal cortex (b) frontal cortex and (c) hippocampus. Animals were treated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035916#pone-0035916-g002" target="_blank">Figure 2</a>. Only vehicle treated animals are shown. N = 5–6/group. * different from Water, +different from TRP+, ♦ different from corresponding group in BALBc mice. 5-HT content was lower at baseline in BALBc in the hippocampus; TRP− significantly decreased 5-HT content in the hippocampus; TRP+ did not consistently increase 5-HT content; TRP− was consistently lower than TRP+ in all regions.</p

Brain 5-HTP.

<p>Brain 5-HTP content in ng/mg tissue in (a) prefrontal cortex (b) frontal cortex and (c) hippocampus. Animals were treated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035916#pone-0035916-g002" target="_blank">Figure 2</a>. Only NSD 1015-treated animals are shown. N = 6/group. * different from Water, +different from TRP+, ♦ different from corresponding group in BALBc mice. TRP− decreased 5-HT synthesis in all brain regions, but the results were only statistically relevant in C57 mice; TRP+ did not increase 5-HT synthesis; at baseline, BALBc mice had a lower level of 5-HT synthesis in the hippocampus and the prefrontal cortex, but not in the frontal cortex.</p

Time Course of Biochemistry Experiment.

<p>Time Course of Biochemistry Experiment.</p

Behavioral data showing time spent in the light.

<p>Only TRP+- and TRP− -treated animals are shown. N = 14/group. +different from TRP+, ♦ different from corresponding group in BALBc mice.</p