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. 2022 Feb 15;4(1):vdac017. doi: 10.1093/noajnl/vdac017

Figure 1.

Figure 1.

IFN-γ increases PD-L1 and IDO1 expression in human glioblastoma cells and extracellular vesicles. (A) Glioblastoma cell lines dBT114 or dBT116 ± 100 ng/mL IFN-γ for 24 h and expression of indicated proteins in whole-cell lysate (WCL) and extracellular vesicles (EVs) was assessed by western blot. (B) Immunoblots from (A) were normalized to loading control (HSP90) by densitometry. Relative intensity compared to baseline (WCL without IFN-γ exposure) is shown (mean ± SEM; n = 3). (C) dBT114 or dBT116 cells ± 10 or 100 ng/mL IFN-γ for 24 h and analyzed by western blot for IDO1, PD-L1, and GAPDH. Bar graphs for PD-L1 show median fluorescence intensity on flow cytometry relative to EVs without IFN-γ exposure (median ± standard deviation, n = 3) while those for IDO1 show relative densitometry intensity compared to baseline (no IFN-γ) after normalization to GAPDH (mean ± standard deviation; n = 3). (D) Nanoparticle tracker analysis histograms and photomicrographs showing the size distribution and frequency of dBT114 and dBT116 EVs ± 100 ng/mL IFN-γ. *P < .05, **P < 0.01,***P < .001, ****P < .0001. IDO1, indoleamine 2,3-dioxygenase 1; IFN-γ, interferon-gamma; PD-L1, programmed cell death ligand 1.