Fig. 1
From: Triggering anti-GBM immune response with EGFR-mediated photoimmunotherapy
ZEGFR:03115-IR700-based PIT in vitro. A Decrease in cell viability as assessed by the CellTiter-Glo® viability assay 24 h post-treatment in GBM cells (U87-MG, U87-MGvIII DKMG, and WSz57) following 1 h incubation with the ZEGFR:03115-IR700 (0.1–0.5 μM) and irradiation with 16 J/cm2 light dose compared to control cells. The results were normalised to the untreated cells (no light, no conjugate) and presented as mean ± SEM (n = 3). Statistical difference in comparison with the control determined using ANOVA with Dunnett’s post hoc test. ****p ≤ 0.0001, ***p ≤ 0.001. B Changes in the percentage of live, apoptotic, and necrotic U87-MGvIII cell populations measured 1, 4, or 24 h post-therapy, following 1 h incubation with ZEGFR:03115-IR700 (0.25 μM) and exposure to 8 or 16 J/cm2 light dose. Data are presented as mean ± SEM (n = 3). Statistical significance in comparison with the control (untreated) was determined using ANOVA with Dunnett’s post hoc test. ****p ≤ 0.0001, ***p ≤ 0.001, *p ≤ 0.05. C Reactive oxygen species (ROS; using DCFDA) production in U87-MGvIII cells, evaluated 10 min post-treatment: 1 h incubation with or without ZEGFR:03115-IR700 (0.25 μM), N-acetyl-l-cysteine (NAC, 5 mM) and irradiation (8 or 16 J/cm2) compared to control cells; 50 μM TBHP was used as a positive control. The results were normalised to the control and presented as mean ± SEM (n = 3). Statistical difference in comparison with the control determined using ANOVA with Dunnett’s post hoc test. ****p ≤ 0.0001, ***p ≤ 0.001. ••p ≤ 0.01 significant decrease in ROS generation after NAC incubation of PIT treated cells. D Decrease in cell viability as assessed by the CellTiter-Glo® viability assay 24 h post-treatment in U87-MGvIII cells, following 1 h incubation with ZEGFR:03115-IR700 (0.1–0.5 μM), with or without N-acetyl-l-cysteine (NAC, 5 mM) and irradiation with 8 or 16 J/cm2 light dose, was confirmed to be dose-dependent and ROS-mediated. Data are presented as mean ± SEM (n = 3). Statistical difference between the groups with or without NAC incubation was determined using the Mann-Whitney test. ****p ≤ 0.0001, *p < 0.05