Representative images of immunohistochemical stains.

<p>(A) Strong PD-L1 staining. (B) Negative PD-L1 staining. (C) PD-L1 positive control. (D) CD8+ high stromal infiltrate. (E) CD8+ tumoral infiltrate. (F) FOXP3 positive stromal infiltrate.</p

Multivariate analysis for OS, DFS.

<p>Multivariate analysis for OS, DFS.</p

PD-L1 and Tumor Infiltrating Lymphocytes as Prognostic Markers in Resected NSCLC

<div><p>Introduction</p><p>Immune checkpoint inhibition has shifted treatment paradigms in non-small cell lung cancer (NSCLC). Conflicting results have been reported regarding the immune infiltrate and programmed death-ligand 1 (PD-L1) as a prognostic marker. We correlated the immune infiltrate and PD-L1 expression with clinicopathologic characteristics in a cohort of resected NSCLC.</p><p>Methods</p><p>A tissue microarray was constructed using triplicate cores from consecutive resected NSCLC. Immunohistochemistry was performed for CD8, FOXP3 and PD-L1. Strong PD-L1 expression was predefined as greater than 50% tumor cell positivity. Matched nodal samples were assessed for concordance of PD-L1 expression.</p><p>Results</p><p>Of 522 patients, 346 were node-negative (N0), 72 N1 and 109 N2; 265 were adenocarcinomas (AC), 182 squamous cell cancers (SCC) and 75 other. Strong PD-L1 expression was found in 24% cases. In the overall cohort, PD-L1 expression was not associated with survival. In patients with N2 disease, strong PD-L1 expression was associated with significantly improved disease-free (DFS) and overall survival (OS) in multivariate analysis (HR 0.49, 95%CI 0.36–0.94, p = 0.031; HR 0.46, 95%CI 0.26–0.80, p = 0.006). In this resected cohort only 5% harboured EGFR mutations, whereas 19% harboured KRAS and 23% other. KRAS mutated tumors were more likely to highly express PD-L1 compared to EGFR (22% vs 3%). A stromal CD8 infiltrate was associated with significantly improved DFS in SCC (HR 0.70, 95%CI 0.50–0.97, p = 0.034), but not AC, whereas FOXP3 was not prognostic. Matched nodal specimens (N = 53) were highly concordant for PD-L1 expression (89%).</p><p>Conclusion</p><p>PD-L1 expression was not prognostic in the overall cohort. PD-L1 expression in primary tumor and matched nodal specimens were highly concordant. The observed survival benefit in N2 disease requires confirmation.</p></div

Reported studies of PD-L1 in NSCLC.

<p>Reported studies of PD-L1 in NSCLC.</p

Additional file 3: Figure S3. of Transketolase-like 1 ectopic expression is associated with DNA hypomethylation and induces the Warburg effect in melanoma cells

TKT and TKTL2 expression in melanoma cells. (A) TKT, (B) TKTL1 and (C) TKTL2 expression in LM-MEL-59 after treatment with two TKTL1 siRNAs or control siRNA for 72 h. Testis tissue was used as positive control for the expression of TKT, TKTL1 and TKTL2. (PPTX 69 kb

Additional file 1: Figure S1. of Transketolase-like 1 ectopic expression is associated with DNA hypomethylation and induces the Warburg effect in melanoma cells

TKTL1 is expressed in a subset of melanoma tumours and melanoma cell lines. (A) Representative staining patterns for TKTL1 in metastatic melanoma tumors are shown. Arrows indicate nuclear staining for Melanin. Original magnification, 200 Οm. (B) QRT-PCR for expression level of TKTL1 in a panel of 53 metastatic melanoma cell lines. (C) High magnification staining pattern of TKTL1 in melanoma cell line LM-MEL-59. Arrows indicate nuclear staining for Melanin. Original magnification, 200 Οm. (PPTX 1827 kb

Additional file 2: Figure S2. of Transketolase-like 1 ectopic expression is associated with DNA hypomethylation and induces the Warburg effect in melanoma cells

Correlation of TKTL1 gene expression with DNA methylation in melanoma patients. Exon expression data was compared to methylation patterns of TKTL1 and Spearman correlation coefficients were calculated. Methylation status at the CpG site (A) cg09892236 and (B) cg23106779 was inversely correlated with TKTL1 gene expression in melanoma samples. (PPTX 114 kb

Additional file 5: Figure S5. of Transketolase-like 1 ectopic expression is associated with DNA hypomethylation and induces the Warburg effect in melanoma cells

TKTL1 expression in melanoma affects proliferation of cells. (A) Measurement of absorbance of LM-MEL-59 cells after TKTL1 siRNA or control siRNA treatment for 48 h by performing MTS assay. (B) Measurement of absorbance of LM-MEL-44 cells after overexpression of TKTL1 or empty vector for 48 h. (PPTX 57 kb

Survival curves demonstrating NY-ESO-1 to be a poor prognostic factor (A) and a predictive marker for benefit from adjuvant chemotherapy (B).

<p>Survival curves demonstrating NY-ESO-1 to be a poor prognostic factor (A) and a predictive marker for benefit from adjuvant chemotherapy (B).</p

Clinicopathological features associated with CTA expression in two cohorts of patients.

<p>Clinicopathological features associated with CTA expression in two cohorts of patients.</p