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. 2017 Oct 25;7:14049. doi: 10.1038/s41598-017-14417-6

Figure 1.

Figure 1

Neuro2a cells express PD-L1 but anti-PD-1 or anti-PD-L1 blocking mAbs have no impact on Neuro2a tumor progression in syngeneic mice. (a) RT-PCR analysis shows that Neuro2apc and NXS2pc cells express PD-L1 mRNA constitutively and after 48 hrs IFN-γ treatment (1,000 IU/ml). Housekeeping gene was β-actin. β-actin gel was cropped from the same gel to uniform the loading order to PD-L1 gel. MW: molecular weight marker 100 bp ladder DNA. CTR-: PCR negative control. (b) Neuro2a and NXS2 cells express surface PD-L1 as detected by immunofluorescence and FACS analysis. Ctr: isotype control PE; baseline: cells stained with anti-PD-L1 PE; induced: cells treated with 1,000 IU/ml rat IFN-γ for 48 hrs and stained with anti-PD-L1 PE. (c) Kaplan-Meier analysis of A/J mice inoculated i.v. with a tumorigenic dose of Neuro2a-luc cells on day 0 and treated with an irrelevant antibody, anti-PD-1, anti-PD-L1 mAb or with anti-PD-1 combined with rIL-21, according to the schedule shown in the inset. Percentages of progression-free mice are indicated on the Y-axis and the fraction of progression-free mice of each group is given in brackets.