Relationship of KLHDC4 expression with clinicopathological characteristics in NPC.

Relationship of KLHDC4 expression with clinicopathological characteristics in NPC.</p

Association between risk of nasopharyngeal carcinoma and seven single-nucleotide polymorphisms.

a<p>Risk allele/reference allele.</p>b<p>OR = odds ratio; CI = confidence interval. OR (95% CI) for each SNP were estimated separately using a logistic regression adjusted for age, sex, educational level, dialect, and rural or urban household type.</p>c<p><i>P</i> values for trend (two-sided) were derived from Cochran- Armitage trend tests.</p

KLHDC4 does not interact with Cullin2 or Cullin3.

(A-B) 293T cells were transiently tranfected with plasmids encoding SFB-tagged KLHDC3, KLHDC4, KLHDC10, KEAP1, or empty vector together with plasmid encoding HA-tagged Cullin2 or Cullin3 as indicated. Cell lysates were precipitated with S-protein beads and immunoblotted with indicated antibodies. (C) Protein structures of KLHDC4, KLHDC3, KLHDC10 and KEAP1. Red rectangle: Kelch repeat; Blue hexagon: BC box; Yellow rhombus: BTB domain; Green rhombus: BACK domain. (TIF)</p

Area under curves (AUC) as a measure of predictive strength for risk-prediction models based on different indicators^a.

a<p>The environmental model is based on consumption of salted fish and preserved vegetables, and cumulative amount of smoking. The family history of NPC model includes family history of NPC only. The epidemiological model combines both environmental and family history of NPC predictors. The genetic risk score model includes a score derived from seven SNPs identified in the Cantonese GWAS. The inclusive model integrates all data on epidemiological and genetic predictors.</p>b<p>χ² statistic and <i>P</i> value was calculated from the Hosmer–Lemeshow Goodness-of-Fit test, a model with χ² statistic <20 (<i>P</i>>0.01) is considered as a good calibration.</p>c<p>AUC of the models were compared with a nonparametric approach, and <i>P</i> value was obtained from the comparison of the inclusive model with the other models.</p

Loss of KLHDC4 reduces NPC cell migration and invasion.

<p>(A) Wound-healing assays were performed at 0 and 20 hours with control and KLHDC4 KO cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of the wound closure area calculated by measuring the decreaseinthe wound bed surface overtime. **<i>P</i> <0.01. (B) KLHDC4 KO significantly reduced the invasive ability of CNE2 cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of average number of cells per field. ***<i>P</i> <0.001.</p

Loss of KLHDC4 induces apoptosis in NPC cells.

<p>(A) Control and KLHDC4 KO cells were stained with DAPI and photographed by fluorescence microscopy under fluorescence and light fields. Left panel: representative images; the arrows indicated the condensed or fragmented nuclear and multiblebbing cells. Right panels: quantification of cells with condensed nuclear per field. **<i>P</i> <0.01. (B) Representative flow cytometric analysis of apoptotic CNE2 control and KLHDC4 KO cells stained for annexin V-FITC/PI under both non-treated and cis-platin treated conditions. The results are summarized in the right panel. Bars denote the S.E.M. (C) Western blot analysis of the expression of cleaved caspase-3, cleaved PARP and GAPDH, as the loading control, under both non-treated and cis-platin treated conditions. (D) Immunohistochemical analysis of KLHDC4, cleaved caspase-3, cleaved PARP, and Ki-67 expression in xenografts generated from CNE2 control and KLHDC4 KO cells. Left panels: representative images; Scale bars: 100 μm. Right panels: quantification of average percentage of cells with positive staining per field; CC3: cleaved caspase-3; CP: cleaved PARP. ***<i>P</i> <0.001.</p

Receiver-operating characteristic (ROC) analysis.

<p>The areas under the ROC curves (AUC) as measures of predictive power for risk-assessment models based on environmental risk factors, family history of NPC, and genetic variants for NPC.</p

KLHDC4 expression correlation in NPC and adjacent nasopharyngeal epithelia.

<p>KLHDC4 expression correlation in NPC and adjacent nasopharyngeal epithelia.</p

Distribution of genetic risk score.

<p>Distribution of the seven SNPs-based genetic risk score in 1,387 NPC cases (black bars) and 1,459 controls (grey bars). Individual risk for NPC was calculated by weighting each risk allele with its corresponding risk coefficient, which was derived from logistic regression.</p

Reclassification of data for use in epidemiological and inclusive models^a.

a<p>NRI: net reclassification improvement; IDI: integrated discrimination index; Reclassification was calculated for strata of predicted risk of <0.2, 0.2 to 0.3, and ≥0.3.</p