MyD88 expression and survival (Kaplan-Meier curves: median survival time shown in months).

<p>MyD88 positive tumours (n = 12) had significantly reduced progression-free survival (A) and overall survival (B) (p = 0.018 and p = 0.008, respectively).</p

TLR4/MyD88 and progression-free survival (Kaplan-Meier curves; median survival shown in months).

<p>TLR4 (A) or MyD88 (B) negative cases had significantly better PFS (15 & 18 months longer; p<0.05).</p

Heterogeneous expression of monoclonal anti-TLR4.

<p>A: variable staining observed in adjacent epithelium within a benign serous cystadenoma (20x). B: focal strong staining within a serous carcinoma (40x).</p

The effect of silencing MyD88 and TLR4 mRNA on the chemoresponsive properties of SKOV-3 cells.

<p>SKOV-3 cells were left untransfected (Unt), transfected with negative control siRNA (siNeg), MyD88 targeting siRNA (siMyD88) or TLR4 targeting siRNA (siTLR4). The transfected cells were incubated for 72 hrs before either harvesting for mRNA analysis (A), for protein analysis (B) or treatment with paclitaxel (C). (A) MyD88 and TLR4 mRNA expression levels were evaluated by TaqMan RT-PCR. MyD88 and TLR4 mRNA expression was normalised to that of an endogenous control, B2M, and calibrated to that of untreated cells to establish the relative percentage of mRNA expression (n = 3, mean +SD). (B) MyD88 and TLR4 mRNA expression levels were evaluated by western blot analysis. GAPDH was used as a loading control. (C) Transfected cells were either left untreated, treated with DMSO (vehicle control) or 3.5 nM of paclitaxel (IC25). 48 hrs post treatment, cell viability was assessed by means of the CCK-8 assay. % cell viability rate was calculated by comparing the absorbance values for the vehicle control to the corresponding paclitaxel treated samples. Results are expressed as mean +SD, n = 3; *p<0.05, **p<0.01 (un-paired Student's t-test).</p

TLR4/MyD88 and overall survival (Kaplan-Meier curves; median survival shown in months).

<p>Survival was longer in MyD88 (B) negative cases (by 19 months; p<0.05). The difference in survival associated with TLR4 (A) is not significant (p>0.5).</p

Quantification of immunohistochemical staining of TLR4 and MyD88 (0 = no staining, 1 = weak staining, 2 = moderate staining, 3 = strong staining).

<p>Quantification of immunohistochemical staining of TLR4 and MyD88 (0 =  no staining, 1 =  weak staining, 2 =  moderate staining, 3 =  strong staining).</p

MiR-21 & miR-146a expression in the test series.

<p>Scatter plots showing relative microRNA expression with standard deviation (fold changes calculated via the 2^−ΔΔCt method). 20 EOC cases (serous carcinomas) grouped as MyD88+ or MyD88- based on protein expression; data shown relative to each group. Average expression of miR-21 & miR-146a increased in MyD88 negative EOC (p<0.05).</p

Distribution of TLR4 & MyD88 protein expression in all patient samples.

<p>Abbreviations: SD, standard deviation; FIGO, Federation International of Gynecology & Obstetrics; NOSE, normal ovarian surface epithelium.</p>†<p>TLR4, MyD88 expression by immunohistochemistry (score >4 =  positive).</p

TLR4 expression pattern.

<p>A: focal weak staining in normal ovarian surface epithelium (40x); B: diffuse moderate staining in a benign serous cystadenoma (20x); C: diffuse strong staining in a borderline serous tumour (10x); D: diffuse strong staining in a (grade 2) serous carcinoma (40x).</p

TLR4 and MyD88 expression in undifferentiated and retinoic acid (RA)-differentiated NTera2 and 2102Ep cells.

<p><b>A</b>, qPCR data: MyD88 expression is down-regulated upon NTera2 differentiation, with minimal changes in TLR4; in contrast 2102Ep cells avoid differentiation and maintain both TLR4 & MyD88 expression (fold changes shown are proportional to internal control gene GAPDH and calculated via the 2^−ΔΔCt method). <b>B</b>, TLR4 and MyD88 protein expression in NTera2 cells: TLR4 staining was unchanged following differentiation (left panel); MyD88 staining in undifferentiated NTera2 cells showed significantly reduced staining after differentiation (right panel) (all 40x). <b>C</b>, TLR4 and MyD88 expression in 2102Ep cells: minimal changes in TLR4 (left panel) or MyD88 (right panel) were observed following RA-treatment (all at 40x).</p