Identification of differentially expressed serum miRNAs with varying doses of radiation at 24 hrs.

<p>A: Heat map generated from the actual counts for 88 miRNAs detected in serum. B: Heat map showing variations in a panel of 18 radio-responsive miRNAs, identified by ANOVA with a cutoff p-value of 0.05. C: Dendrogram with a panel of markers identified with coefficient of variance across samples with a cutoff value of 0.4. D: Overlapping set of miRNAs from ANOVA and CV. Red marks high and green marks low expression.</p

Dose and time dependent depletion of miRNA-150 in animals exposed to 1, 2, 4, 6 and 8 Gy with reference to controls analyzed at 24 hrs (A) and 48 hrs (B).

<p>Statistical analysis was performed using an unpaired two-tailed students t-test (*) = <i>p<0.05</i>; (**) = <i>p<0.005;</i> (***) = <i>p<0.0005</i>. C: Kinetics of depletion of miRNA-150 as a function of dose and time relative to respective controls.</p

Variations in the counts of selected miRNAs following fractionated radiation.

<p>The fluorescent counts obtained after normalization were plotted for individual animals with the dose and time as given. A: The dose dependent depletion of serum miRNA-150 at different time points during and after fractionation (p-values: 4 Gy- 0.0003, 8 Gy- 0.0001, 12 Gy- 0.0001). B: Counts from a non-responsive molecule miRNA-23a (control). C and D: Radiation induced increases in miRNA-200b (p-values 4 Gy- 0.014, 8 Gy- 0.0047, 12 Gy 0.0027) and miRNA-762.</p

Analysis of fold variations of selected serum miRNA biomarkers with a clear dose response.

<p>Histograms show variations in the fluorescent counts detected in the nCounter multiplex assay, plotted against treatment. The counts obtained after normalization using multiple spike-in oligos were plotted for individual animals. A: The dose dependent depletion of serum miRNA-150 at 24 hrs (p-values: 1 Gy- 0.0164, 2 Gy- 0.0191, 4 Gy- 0.0026, 6 Gy- 0.0001, 8 Gy- 0.0001). B: Counts from a non-responsive molecule miRNA-23a, comparable to that of miRNA-150 in control animals. C and D: Radiation induced increase in miRNA-200b and miRNA-762 (p-values, miRNA-200b:1 Gy- 0.7172, 2 Gy-0.4193, 4 Gy- 0.4231, 6 Gy- 0.0421, 8 Gy- 0.0296; miRNA-762∶1 Gy- 0.4061, 2 Gy- 0.1675, 4 Gy- 0.0324, 6 Gy- 0.3139, 8 Gy- 0.001).</p

Identification of miRNAs derived from red blood cells that potentially contaminate serum samples due to increased levels of hemolysis.

<p>The X- axis shows samples with increasing levels (+) of hemolysis and the Y-axis shows the actual counts of several miRNAs that are altered with an increase in hemolysis.</p

A list of miRNAs detected in mouse serum, arranged in their order of abundance.

<p>The average signals (counts) detected from a total of 20 µl serum RNA preparations from three different control mice belonging to two different mice strains (A: CBA/J and B: C57BL/6) are presented. The miRNAs are listed in the order of abundance.</p