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. 2021 Jan 28;11:609490. doi: 10.3389/fimmu.2020.609490

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Copyright © 2021 Grobben, de Man, van Doornmalen, Muller, Willemsen-Seegers, Vu-Pham, Mulder, Prinsen, de Wit, Sterrenburg, van Cauter, den Ouden, van Altena, Massuger, Uitdehaag, Buijsman and Zaman

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Development of a syngeneic mouse model using a B16F10 melanoma cell line stably overexpressing mIDO1. (A) Analysis of mIDO1 mRNA levels by qPCR in the parental B16F10 cell line and two mIDO1-transfected stable sublines (i6 and j19). mRNA levels were normalized for the expression of three housekeeping genes (ACTB, GAPDH, and RPL37), and scaled based on the hepatoma Hepa 1-6 cell line stimulated with IFNγ (included as a positive control). (B) Immunoblot analysis of the parental B16F10 cell line and mIDO1-overexpressing sublines. (C) Mean tumor volume after implantation of the B16F10-mIDO1 i6 and j19 sublines in B6D2F1 mice. Results are expressed as mean ± SEM. (D) Analysis of mIDO1 mRNA levels in the tumor tissues by qPCR. mRNA levels are scaled based on the mRNA level of the B16F10-mIDO1 subline j19 in cell culture. (E) Intratumoral Trp and (F) Kyn levels as determined by LC-MS/MS. LLOQ in (E, F) indicates the lower limit of quantification of the experiment. Results of (D–F) are expressed as mean ± SD.