viRNA coverage of the LACV/Human/1960 strain genome in C6/36 and S2 cells.

<p>Complete genome of LACV/Human/1960 strain showing intensity at each nucleotide of the genome in C6/36 (A,C,E) and S2 (B,D,F) cells. A and B correspond with the L gene segment (6,980 nt), C and D the M gene segment (4,526 nt), and E and F to the S gene segment (984 nt). Plotted are the 19–30-mer viRNA reads across the length of each segment represented by the x-axis. Reads originating from the genomic, negative strand are represented in red below the x-axis and those originating from the positive strand are represented in blue above the x-axis.</p

Size and abundance of small RNA reads Mmapping to the viral genomes.

<p>The abundance of 19–30-mer sRNA reads mapping to the WNV (A), SINV (B) and LACV (C) genomes based on size. Abundance is represented as a percentage of the total viRNAs from each sample. The black bars correspond with samples collected from S2 cells and white bars from C6/36 cells.</p

viRNA coverage of the TE3′2J SINV genome in C6/36 and S2 cells.

<p>Complete genome of TE3′2J SINV (11,385 nt.) showing intensity at each nucleotide of the genome in C6/36 (A) and S2 (B) cells. Plotted are the 19–30-mer viRNA reads. Reads originating from the genomic, positive strand are represented in blue above the x-axis and those originating from the negative strand are represented in red below the x-axis. The green vertical line represents the location of the subgenomic promoter.</p

Small RNA profiles from C6/36 and S2 infected cells.

<p>Small RNA profiles from C6/36 and S2 infected cells.</p

Pathway results for Danish sister pairs (n = 93); showing only genes related to the Discovery findings.

<p>Pathway results for Danish sister pairs (n = 93); showing only genes related to the Discovery findings.</p

Most significant pathway results for Finnish mothers with recurrent SPTB (n = 10).

<p>Most significant pathway results for Finnish mothers with recurrent SPTB (n = 10).</p

<i>HSPA1L</i> variants in the large GWAS data and their association to SPTB.

<p><i>HSPA1L</i> variants in the large GWAS data and their association to SPTB.</p

Effects of <i>HSPA1L</i> variants on protein sequence and structure.

<p>(A) HSPA1L is a 641-amino acid protein that consist of two major functional domains; an N-terminal nucleotide-binding domain and a C-terminal substrate-binding domain, that are connected with interdomain linker. Locations of four <i>HSPA1L</i> variants from whole exome sequencing (Discovery and Replication findings) are shown in the protein sequence. Purple diamonds represent ATP nucleotide-binding sites at positions 14−17, 204−206, 270−277 and 341−344. Green circles denote the additional phosphorylation site generated by Ala268Thr and the existing one T267-p. (B) <i>In silico</i> comparison of higher order assembly of reference and modified (Ala268Thr) HSPA1L protein models containing an ADP molecule. Overlayed reference and Ala268Thr molecules are presented as gold and light blue rounded ribbon structures, respectively. The interacting ADP molecule is shown as a stick model. (C) Closeup view of the intermolecular contact interface of HSPA1L bound to the ADP molecule. Key interacting residues (amino acids) are shown and their corresponding side chains are presented as stick molecules; nitrogen and oxygen atoms are indicated in blue and red, respectively. All interacting residues; THR16, TYR17, GLU270, LYS273, ARG274, SER277 (not shown in figure) and ASP368, that bind to the ADP ligand showed small changes in the chemical bond lengths. Only changes of ≥0.002Å are shown in the figure (+/- lenght of Ala268Thr structure in relation to reference structure).</p

Overview of the whole exome sequencing study workflow.

<p>Abbreviations used QC; quality control, MAF; minor allele frequency, RD; read depth, GQ; genotype quality.</p

HSPA1L and GR protein levels in decidualized human endometrial stromal fibroblasts.

<p>Cultured ESFs were transfected with WT or Ala268Thr <i>HSPA1L</i>-pcDNA3.1 constructs or with empty pcDNA3.1 vector (control). Cells were treated with decidualization media supplemented with 100nM dexamethasone (glucocorticoids) for 72h. Both cytosolic and nuclear protein were extracted, and HSPA1L and GR protein levels were measured by Western blot. Band intensity of HSPA1L or GR was normalized to band intensity of the corresponding β-actin. Cytosolic (A) and nuclear (B) HSPA1L levels as well as cytosolic (C) and nuclear (D) GR levels are shown for control (empty vector), WT and Ala268Thr sample groups. Each experiment was performed as triplicates in three different passages (n = 9 each group, except n = 8 for nuclear control group) and bars represent mean + SEM. Significant p-value <0.05 is presented with an asterisk. Cytosolic GR levels were significantly higher (p = 0.04) in the WT compared to the Ala268Thr group as well as in the WT compared to the control group (p = 0.04).</p