Characteristics of the 31 selected genes.

<p>Characteristics of the 31 selected genes.</p

Large-scale studies assessing gene expression changes in mouse blood after irradiation.

<p>Large-scale studies assessing gene expression changes in mouse blood after irradiation.</p

Estimated specificity, positive predictive value rate, false positive and Youden’s index of the 33 genes discriminating radiation dose <2Gy and ≥2Gy at fixed sensitivity of 100%.

<p>Estimated specificity, positive predictive value rate, false positive and Youden’s index of the 33 genes discriminating radiation dose <2Gy and ≥2Gy at fixed sensitivity of 100%.</p

Receiver operating characteristic (ROC) curve analysis of IER5 + TNFSF4 combination that displays area under the ROC curve (AUC) ≥ 0.99 to discriminate radiation dose < 2 Gy from radiation dose ≥ 2 Gy.

<p>Receiver operating characteristic (ROC) curve analysis of IER5 + TNFSF4 combination that displays area under the ROC curve (AUC) ≥ 0.99 to discriminate radiation dose < 2 Gy from radiation dose ≥ 2 Gy.</p

AUC and diagnostic accuracies of the 33 genes discriminating radiation dose <2Gy and ≥2Gy classified according to their maximized Youden’s index.

<p>AUC and diagnostic accuracies of the 33 genes discriminating radiation dose <2Gy and ≥2Gy classified according to their maximized Youden’s index.</p

Large-scale studies assessing gene expression changes in human blood after particle irradiation.

<p>Large-scale studies assessing gene expression changes in human blood after particle irradiation.</p

Flow diagram outlining the selection procedure to identify 24 articles that were included in the systematic review of gene radiation dosimetry biomarkers in human blood.

<p>Flow diagram outlining the selection procedure to identify 24 articles that were included in the systematic review of gene radiation dosimetry biomarkers in human blood.</p

sCLU expression is increased during senescence.

<p>Young (Y), middle-age (M), premature-senescent (PS), senescent (S), and late senescent (LS) IMR-90 cells were generated by continuous culture. (A) The “age” of the IMR-90 cells was determined by population doublings and %SA-β-gal+ cell measurements. Experiments were repeated three or more times in triplicate each, and representative results are shown. (PD, population doubling). (B) Senescence markers, phospho-ser15-p53, total p53, p21 and p16 levels, were markedly increased during senescence. (C) Both the precursor (psCLU) and the mature secretory (sCLU) forms of clusterin were induced in total cell lysates during senescence. sCLU levels increased in the media of senescent IMR-90 cells. (D) The promoter activity of sCLU was increased during senescence. *, p-value≤0.05, **, p-value≤0.01.</p

sCLU induction during senescence is mediated through IGF-1R/MAPK/Erk/Egr-1 signaling pathway.

<p>(A) IGF-1 expression was increased in senescent IMR-90 cells. IGF-1 levels in media from young or senescent IMR-90 cells were measured using ELISAs as described above, and normalized to media volume and adjusted cell number (4×10⁴ cells). (B) Total IGF-1 receptor (IGF-1R), phosphorylated IGF-1 receptor (p-IGF-1R), Src, phosphorylated Src, Erk-1/2, phosphorylated-Erk-1/2, and Egr-1 levels in young (Y), middle-age (M), premature-senescent (PS), and senescent (S) IMR-90 cells were determined by western blot analyses. β-Actin levels were measured to monitor loading. (C) Exposure to the indicated doses of the IGF-1R tyrosine kinase inhibitor (AG1024) repressed sCLU levels in senescent IMR-90 cells. psCLU and sCLU were detected by western blot analysis. β-Actin levels were monitored to show equal loading. Senescent IMR-90 cells exposed to varying doses of AG1024 (µM as indicated for 24 h) showed dose-dependent increases in apoptosis (monitored by TUNEL + staining). Only senescent, and not young or middle-aged IMR-90, cells were hypersensitive to AG1024. In the experiment outlined below the western blot analyses cells were exposed to AG1024 (5 µM) for 48 h and therefore higher levels of apoptotic cells were noted in senescent IMR-90 cells compared to cells exposed for 24 h above. Note that only senescent cells were hypersensitive to these inhibitor treatments. All experiments were repeated three times in triplicate.</p

sCLU protects cells from senescence.

<p>(A) sCLU was knocked down in an IMR-90 pooled population containing a lentiviral-mediated inducible shRNA-sCLU construct cultured in presence of doxycycline (dox), while expression of sCLU was not affect in the same pooled population in the absence of doxycycline (dox). The population doubling of IMR-90 cells with inducible shRNA-sCLU decreased faster when cultured in the presence of dox compared to in cells cultured without dox. **, p-value of two curves was ≤0.01. Experiments were repeated three times, starting from infection of young IMR-90 cells with selection of a pTRIPZ-shCLU-containing pooled population. Similar results were observed in all three experiments and representative results from one experiment are shown. (B) IMR-90 cells with sCLU knockdown had significantly higher %SA-β-gal+ cells after being cultured for 33, 39, and 53 days with or without dox. SA-β-gal+ staining was performed to identify senescent cells. *, p-value ≤0.05. (C) Controls included a pooled IMR-90 cell population contain an inducible shRNA-non-targeted, scrambled shRNA construct were cultured in presence or absence of dox. Growth rates and sCLU levels were not affected in the shRNA-Scr (non-targeted) IMR-90 cell populations. Population doubling of IMR-90 cells with inducible non-targeting shRNA was not different when cultured with/without doxycycline. The experiments were repeated three times similarly as inducible shCLU experiments. Representative results of three experiments were shown here. (D) After culturing for 38, 45, and 65 days with or without dox, SA-β-gal+ staining was performed to identify senescent cells in IMR-90 pooled populations with inducible shRNA-Scr (non-targeting) vector. (E) Specificity of shCLU knockdown in conditional IMR-90 senescent cells. IMR-90 pooled populations containing the pTRIPZ-shCLU vector was cultured with or without doxcycline (100 nM for 3 days) and sCLU and nCLU levels were monitored using Western blot analyses. α-Tubulin levels were shown as a loading control. Note that sCLU and not nCLU levels were specifically knocked down (>90%) in cells exposed to dox. In these experiments, IMR-90 cells were cultured in 95% air-5% CO₂, estimated at ∼20% O₂.</p