Time-dependent CK1δ and CK1ε proteomics.

<p>(<b>A</b>) Illustration of screen. Cells stably expressing dually-tagged (yellow hexagon) CK1 were treated with dexamethasone (DEX) and harvested at different time points. Protein complexes were stabilized with DSP prior to lysis and purification for peptide identification by LC-MS/MS. (<b>B</b>) HA immunoblot showing DSP efficacy and the effect of dexamethasone on CK1ε complexes. (<b>C</b>) Diagram showing proteins pulled out at the different time points by CK1δ and CK1ε. Numbers represent hours after DEX that cells were harvested. Open circles were pulled out with both CK1δ and CK1ε (orange circles are known circadian proteins). Closed circles were pulled out with CK1δ alone (black-fill) or CK1ε alone (red-fill).</p

PHB2 is molecular clock component.

<p>(<b>A</b>) Representative graph (n = 3) of M34-luciferase reporter assays following transient cotransfections of M34-luc, renilla, GFP and indicated siRNAs without (left) or with (right) <i>BMAL1</i> and <i>CLOCK</i>. (<b>B</b>) Representative graph (n = 3) showing transcript levels of <i>PER2</i> and <i>PHB2</i> 24 h after transfecting cells with control, <i>PHB2</i>, <i>CK1δ</i>, <i>CK1ε</i> and <i>PER2</i> siRNA. (<b>C</b>) Graph (n = 3) showing the effect of indicated siRNAs without (left, marked as control) or with (right, marked as <i>PHB2</i>) <i>PHB2</i> siRNA on PHB2 protein levels relative to ACTIN. (<b>D</b>) Illustration summarizing molecular mechanism based on our results where shapes indicate proteins. (* indicates p<0.05; ** indicates p<0.01; determined by student t-test analysis. Error bars indicate STDEV).</p

Cell-based kinase and Lumicycle assays.

<p>(<b>A</b>) Autoradiography (P³²) and immunoblots of HA-tagged proteins (green) and GFP-tagged CK1δ or CK1ε (red). Graph <i>inset</i> showing percent change of SAPS3 phosphorylation by CK1δ and CK1ε (n = 2). (<b>B</b>) Results from LumiCycle assays in cells transfected with indicated siRNAs. (<b>C</b>) Representative traces after cell synchronization of control (gray), <i>CRY2</i> (green) and <i>PHB2</i> (white) siRNAs (<b><i>left</i></b>). Graph showing effects of control siRNA, <i>CRY2</i> siRNA, two <i>PHB2</i> siRNAs (<i>PHB</i>2.1 & <i>PHB</i>2.2) and three <i>PHB1</i> siRNAs (<i>PHB</i>1.1, <i>PHB</i>1.2, <i>PHB</i>1.3) (<i>right</i>). (Sample size indicated by number inside each bar, ** indicates p<0.001 following regression analysis. Error bars indicate SEM).</p

Correction: Alternating Hemiplegia of Childhood: Retrospective Genetic Study and Genotype-Phenotype Correlations in 187 Subjects from the US AHCF Registry

<p>Correction: Alternating Hemiplegia of Childhood: Retrospective Genetic Study and Genotype-Phenotype Correlations in 187 Subjects from the US AHCF Registry</p

Schematic representation of <i>ATP1A3</i> mutations.

<p>Mutations identified in our cohort are indicated above the gene; all the mutations previously published are indicated in black; novel mutations are indicated in light blue; mutations identified in multiplex cases are underlined; mutations reported in DYT12 are indicated in green; the mutation reported in CAPOS syndrome is indicated in red. The mutation associated with a phenotype combining features of both AHC and RDP is in orange. The 2 most common mutations are in bold. Asterisks mean that 2 different nucleotide changes have been identified for these protein variants.</p

Ages at unsupported sitting acquisition in each group of patients defined by their genotype.

<p>Cumulative probability of acquiring unsupported sitting by patients presenting the E815K mutation, compared to patientsmutation (3b). Patients with the E815K mutation are likely to gain unsupported sitting at a later age than patients in each of the other groups (respectively P = 0.0002 and P = 0.0020).</p

Summary of the 187 patients included in the genetic study.

<p>Summary of the 187 patients included in the genetic study.</p

Ages at onset of AHC in each group of patients defined by their genotype.

<p>The horizontal lines in the boxes indicate the 25th percentile (bottom), the median (middle) and the 75 percentile (top) values. Crosses indicate the mean values. Numbers of patients analyzed in each group are indicated above the boxes.</p

Odds ratio for occurrence of status epilepticus in AHC patients with different <i>ATP1A3</i> mutations.

<p>Odds ratio for occurrence of status epilepticus in AHC patients with different <i>ATP1A3</i> mutations.</p

Summary of 164 AHC patients included in the genotype-phenotype correlation study.

<p>Summary of 164 AHC patients included in the genotype-phenotype correlation study.</p