Fig. 3
From: Antitumor effects of a novel glucose-conjugated bacteriochlorin for photodynamic therapy
Evaluation of cellular uptake, subcellular localization, ROS and singlet oxygen generation, and apoptosis induction of photosensitizers. (a) Flow cytometric histograms showing time-dependent uptake of Glc-TFPB in HCT116 cells. The x-axis represents fluorescence intensity, and the y-axis represents the number of cells. (b) Quantification of cellular uptake expressed as mean fluorescence intensity (MFI). Data are presented as the mean ± SEM (n = 3). (c) Confocal microscopy images showing subcellular localization of Glc-TFPB (10 µM, 24 h) in HCT116 cells stained with organelle-specific fluorescent probes. Original magnification × 300; scale bar 10 µm. (d) Quantitative analysis of subcellular localization of Glc-TFPB. Data are represented as mean ± SEM (n = 6 per organelle). (e) Comparison of ROS generation between Glc-TFPB-PDT and TS-PDT. HCT116 cells were incubated with 5 µM Glc-TFPB or TS for 24 h, then either irradiated with light or left unirradiated, stained with H₂DFFDA, and analyzed by flow cytometry. Representative data from one of three independent experiments are shown. (f) Quantification of ROS generation based on MFI under different conditions (TS or Glc-TFPB treatment with or without PDT). Data are presented as mean ± SEM (n = 3). (g) Singlet oxygen generation detected using SOSG at selected concentrations of Glc-TFPB after light irradiation. Representative fluorescence spectra from one of three independent experiments are shown. (h) Flow cytometric analysis of apoptosis based on active caspase-3 levels after Glc-TFPB-PDT. HCT116 cells were incubated with 4 µM Glc-TFPB, with or without light irradiation, stained with the PE Active Caspase-3 Apoptosis Kit, and analyzed by flow cytometry. Representative histograms from one of three independent experiments are shown. (i) Quantification of active caspase-3 expression based on MFI. Data are presented as mean ± SEM (n = 3). Statistical analyses were performed using the Holm–Sidak test for (d) and Tukey’s multiple comparisons test for (f) and (i). Differences were considered statistically significant at ***p < 0.001.
