Fig. 2
From: Triggering anti-GBM immune response with EGFR-mediated photoimmunotherapy
Immunogenic cell death (ICD) of GBM cells following EGFR-targeted PIT. A, B Concentrations of ATP and HMGB1 proteins released into the medium from U87-MGvIII cells over time (5, 30 min; 1, 4, or 24 h) post-treatment (1 h incubation with or without ZEGFR:03115-IR700 (0.25 μM) and irradiation (16 J/cm2)). Data are presented as mean ± SEM (n = 3). Statistical significance in comparison with the control (untreated) group was determined using ANOVA with Dunnett’s post hoc test. ***p ≤ 0.001. C Western blot assessment of EGFR, HSP70, HSP90, calreticulin (CRT) and HMGB1 expression levels in U87-MGvIII cells and cell supernatants (medium) over time (5 min; 1, 4, 8, or 24 h) post-treatment (PIT: 0.25 μM ZEGFR:03115-IR700 + 16 J/cm2) in comparison with irradiated only (16 J/cm2) and control cells. β-Actin was used as a loading control. D, F DC maturation after co-culturing with ZEGFR:03115-IR700-treated (1 h incubation, 0.25 μM) U87-MGvIII cells post-irradiation with 16 J/cm2 light dose. CD86, HLA-DR, and CD40 expression level on the surface of DC cell membrane (live, CD14-negative and CD11c-positive population) as measured by flow cytometry 48 h post-treatment. Immature DCs (iDC) cultured without stimulation for maturation were used as a control. E. coli lipopolysaccharide (LPS)-treated iDC were used as a positive control. Data are presented as mean ± SEM (n = 3–4). The graphs represent the data from four healthy blood donors. Statistical significance in comparison with the iDC group was determined using ANOVA with the Holm-Sidak correction test. **p ≤ 0.01, *p ≤ 0.05