Article Figures & Data
Figures
- Figure 1.
SOX10 increased the expression of PD-L1 in A375 cells. (A) Expression of SOX10 in SOX10-knockdown cells. A375 cells were transfected with SOX10 siRNA, and 48 h later, the cells were treated with IFNγ at several concentrations for 12 h, after which total RNA was obtained and the expression of SOX10 was assessed by real-time quantitative RT-PCR. Data are shown as means±SEM. (B) Expression of SOX10 in SOX10-overexpressed cells. A375 cells were transfected with SOX10 plasmid, and 48 h later, the cells were treated with IFNγ at several concentrations for 12 h, after which total RNA was obtained and the expression of SOX10 was assessed by real-time quantitative RT-PCR. Data are shown as means±SEM. The expression of levels of SOX10 showed no significant difference by IFNγ treatment. (C) Expression of PD-L1 in SOX10-knockdown and -overexpressing cells. A375 cells were transfected with SOX10 siRNA or SOX10 plasmid, and 48 h later, the cells were treated with IFNγ at several concentrations for 12 h, after which total RNA was obtained and the expression of PD-L1 was assessed by real-time quantitative RT-PCR. Data are shown as means±SEM. SOX10 knockdown (KD) significantly decreased the expression of PD-L1, on the other hand SOX10 overexpression (OE) significantly increased the expression of PD-L1. (D) PD-L1 expression under SOX10 knockdown and overexpression in 888-mel cells and SK-MEL-28 cells. 888-mel cells and SK-MEL-28 cells were transfected with SOX10 siRNA or SOX10 plasmid, 2 days later the expression of PD-L1 was addressed by qRT-PCR. Data are shown as mean±SEM. SOX10 knockdown (KD) significantly decreased the expression of PD-L1, on the other hand SOX10 overexpression (OE) significantly increased the expression of PD-L1. CTRL: Control.
- Figure 2.
PD-L1 expression in SOX10 overexpression and knockdown. A375 cells were transfected with SOX10 siRNA or SOX10 plasmid, and 48 h later, the cells were treated with IFNγ at several concentrations for 12 h, after which the expression of PD-L1 was assessed by FACS. Data are shown as means±SEM. SOX10 knockdown (KD) decreased the expression of PD-L1, on the other hand SOX10 overexpression (OE) significantly increased the expression of PD-L1 at protein levels. CTRL: Control.
- Figure 3.
Western blots of PD-L1 and IRF-1 under SOX10 overexpression. A375 cells were transfected with SOX10 plasmid, and 48 h later, the cells were treated with IFNγ at several concentrations for 12 h, after which the expression of IRF-1 and PD-L1 were assessed by western blotting. β-actin was used as an internal positive control. The numerical data indicate the relative quantified data of the bands. SOX10 overexpression (OE) increased the expression of PD-L1 and upstream protein IRF-1 at protein levels.
- Figure 4.
Overexpression of SOX10 enabled melanoma cell escape from TCR-T cells. (A) IFNγ ELISPOT assay of TCR-T cells. NY-ESO-1157-165–specific TCR-transduced TCR-T cells were used. T2 cells were pulsed with NY-ESO-1157-165 peptide and used for IFNγ ELISPOT assay. HIV peptide pulsed (+) and un-pulsed (−) samples were used as negative controls. (B) Expression of PD-1 in TCR-T cells treated with CD3/CD28 beads. NY-ESO-1157-165–specific TCR-T cells were treated with CD3/CD28 beads for 3 days, and the expression of PD-1 was evaluated by FACS. An un-treated sample (CD3/CD28 beads−) was used as a negative control. Isotype antibody was used a negative control for FACS. (C) IFNγ ELISPOT assay of PD-1+ TCR-T cells for SOX10-overexpressed A375 cells. SOX10-overexpressed A375 cells were used for IFNγ ELISPOT assay. CD3/CD28 beads+ TCR-T cells were used as effector cells. CD3/CD28 beads− TCR-T cells were used as a negative control. Data are shown as means±SEM. Statistical significance was evaluated by the Student’s t-test. SOX10 overexpression decreased the TCR-T reaction of CD3/CD28 beads treated PD-1-positive cells, whereas did not decrease the CD3/CD28 bead un-treated PD-1-negative cells.
