Interleukin-10 regulated tumour tolerance in non-small cell lung cancer

Julius Malte Vahl1&, Juliane Friedrich1, Susanne Mittler1, Sonja Trump1, Lisanne Heim1, Katerina Kachler1, Liubov Balabko1, Nicole Fuhrich2, Carol-Immanuel Geppert2, Denis Iulian Trufa3, Nina Sopel1, Ralf Rieker2, Horia Sirbu3 and Susetta Finotto1**

1 Department ofMolecularPneumology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Erlangen, Germany.

2 Institute ofPathology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Erlangen, Germany

3Department of Thoracic Surgery, Friedrich-Alexander-UniversitätErlangen-Nürnberg(FAU), Erlangen, Germany.

The present work was performed in fulfillment of the requirements for obtaining the degree “Dr. med.’’for Julius Vahl

Running Title: IL-10 and NSCLC

Financial Support: this work was supported by an IZKF grant number A59 in Erlangen. Julius M. Vahl received a fellowship from the SFB 643 on Cellular Immune-intervention in Erlangen.

**Corresponding author

Prof. Susetta Finotto, PhD

Laboratories of Cellular and Molecular Lung Immunology

Department ofMolecularPneumology

Friedrich-Alexander-Universität Erlangen-Nürnberg

Hartmannstraße 14

91052 Erlangen, Germany

Phone: 0049-9131-8542454

Email:

Conflict of Interest

The authors declare no conflict of interest on the matter described in this manuscript

Supplementary Figure 1:Negative controls of the different used immunohistochemicalstainings and the calculation of the area fraction with Image J

(A) Negative control for Figure 1A was performed in non-small cell lung cancer tumour and control sector. (B) Negative control for Figure 1C and D was performed in a control block including different types of tissue and lung tissue (of which a cutout is shown). (C) Negative control for Figure 2A and B was performed in non-small cell lung cancer tumour and control sector. (D) Negative control for Figure 3A and B was performed in a control block including different types of tissue and lung tissue (cutout is shown). Concerning the negative control of Figure 5A, we refer to negative control of Figure 2 and 3 (E) Explanation of the area fraction-calculation with Image J. The area fraction is the quotient between the marked area of interest (dividend [pixel]), respectively all positively immune-histochemical stained cells and nearly all cell material of an image forming a second area (divisor [pixel]) excluding interspace areas. The required area selections to build this quotient are performed via specific settings in color threshold of Image J. If this dimensionless quotient is multiplied by 100, the percentage of stained cells in the investigated picture is obtained.