Table 1: Autism Diagnostic Interview-Revised (ADI-R) algorithm domain scores Identifier Sex Diagnosis Y.O.B. Social (cutoff=10) Communication (cutoff=8) Repetitive (cutoff=3) Evident before 36 Verbal months AU078103 Female Spectrum 1993 9 6 6 yes yes AU078104 Male Autism 1995 20 22 8 yes yes AU078105 Male Autism 1997 21 15 7 yes yes Simons Simplex Collection 13344.p1 Male Autism 1998 24 14 11 yes yes Simons Simplex Collection 13344.s1 Female Designated sibling 2000 N/A N/A N/A N/A N/A ASD collection Autism Sequencing Consortium Autism Sequencing Consortium Autism Sequencing Consortium Supplemental Table 1:. Autism Diagnostic Interview-Revised (ADI-R) algorithm domain scores. Trained clinicians conducted the ADI-R and algorithm scores were calculated. ADIR subdomain scores with diagnostic cutoffs are shown along with the autism cohort, subject identifier, sex, year of birth and verbal capabilities of the individuals. Individuals received a diagnosis of “autism” or “spectrum” based on measures from the ADIR and Autism Diagnostic Observation Schedule (ADOS). DAT STX1 Merge STX1A STX1B p-STX1 5 µm Supplemental Figure 1: SXT1A, STX1B and p-STX1 colocalize with DAT in the striatum. Striatal slices were reacted with anti-DAT antibodies (green) and the appropriate antiSTX1 antibodies (red) as indicated, processed, and imaged by confocal microscopy. A SCG neurons presynaptic marker: NET p-STX1 merge 20 µm 1 µm 20 µm B SCG neurons vehicle p-STX1 AMPH C SCG neurons STX1 CK2α DIC 5 µm merge NET 5 µm p-STX1/NET (% control) 200 150 100 D striatum STX1 DAT merge DAT merge * 5 µm 50 p-STX1 0 5 µm Supplemental Figure 2: AMPH treatment increases p-STX1 levels in boutons: (A) Mouse superior cervical ganglion (SCG) neurons were fixed and p-STX1 (green) was visualized by immunoflourescence. The norepinephrine transporter (NET, red) was used as a marker of presynaptic boutons. The merge shows that p-STX1 is present in presynaptic boutons of cultured catecholamine neurons. Inset: Confocal section of a single presynaptic bouton (arrow). (B) Top: SCG neurons were treated with either vehicle or 10 µM AMPH for 20 minutes. Cells were fixed and immunostained for p-STX1 (green) and for the NET (red). AMPH increases the intensity of p-STX1 in presynaptic boutons. Bottom: p-STX1 levels were quantified by the ratio of p-STX1 pixel intensity normalized to NET pixel intensity. AMPH significantly increases the intensity of p-STX1 in presynaptic boutons (* = p < 0.05 by Student’s t-test; n = 5-6). (C) In SCG neurons cultured from NET KO mice, p-Stx1 colocalizes with CK2. (D) In mouse striatal slices, DAT colocalizes with STX1 and p-STX1.
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