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Fig. 4 | Journal for ImmunoTherapy of Cancer

Fig. 4

From: Immunogenic cell death induced by a new photodynamic therapy based on photosens and photodithazine

Fig. 4

Phagocytosis assay and analysis of BMDCs maturation in vitro. Tumor cells dying after treatment with PS-PDT or PD-PDT were efficiently engulfed by BMDCs in vitro (a and d). The data for the uptake of GL261 (a) and MCA205 (d) cells treated with PS-PDT or PD-PDT represent the mean values ± SEM of the duplicates from three independent experiments The rate of phagocytosis increased with the increase in the number of dying/dead cells (1:1 versus 1:5). Statistical significance was calculated by two-way ANOVA, *p < 0.01. Representative flow cytometry dot plots show the uptake of CMFDA-labeled dead GL261 and MCA205 cell material by BMDCs (CD11c+CMFDA+ double-positive cells) are shown in the Additional file 3: Figure S3A, S3B. b-f Tumor cells dying after PS-PDT or PD-PDT treatment induce BMDC maturation in vitro. Co-culture of BMDCs with dying GL261 (b) and MCA205 (e) cells in two different ratios (1:1 and 1:5) and the percentage of CD11c+CD86+ BMDCs is expressed as the mean value ±SEM. Statistical significance was calculated by a Mann-Whitney non parametric t-test, *p < 0.01. Co-culture of BMDCs with dying GL261 (c) and MCA205 (f) cells after treatment with PS-PDT or PD-PDT in two different ratios (1:1 and 1:5) and the percentage of CD11c+CD40+ BMDCs is expressed as the mean value ±SEM of five independent experiments for PS-PDT and four independent experiments for PD-PDT; each experiment was done in the duplicate. In all figure panels, BMDCs stimulated with LPS served as a positive control. MCA205 cells subjected to the several rounds of freeze-thaw (F/T) cycles were used as a negative control in (e and f). Statistical significance was calculated by a Mann-Whitney non parametric t-test, p < 0.05. g Absolute concentrations of IL-6 are the mean values ± SEM from three independent experiments in the co-cultures of BMDCs with the respective target MCA205 cells at three different ratios (1:1, 1:5 and 1:10). LPS treated BMDCs were used as a positive control. Statistical significance was calculated by a Mann-Whitney non parametric t-test. The differences are shown by comparing the respective group with BMDCs co-cultured with either *live MCA205 or #F/T MCA205 cells. p < 0.03

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