Figures & data
Figure 1 Schematic formation of 5-FU-loaded BSA-LTB nanoparticles.
Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.
![Figure 1 Schematic formation of 5-FU-loaded BSA-LTB nanoparticles.Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.](/cms/asset/edadbe85-2474-4a8d-abd1-d4e2cbce8a3f/dijn_a_60764_f0001_c.jpg)
Figure 2 Fluorescence microscope images of identification of LTB with immunofluorescence.
Notes: (A) BSA-LTB nanoparticles; (B) BSA nanoparticles.
Abbreviations: BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.
![Figure 2 Fluorescence microscope images of identification of LTB with immunofluorescence.Notes: (A) BSA-LTB nanoparticles; (B) BSA nanoparticles.Abbreviations: BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.](/cms/asset/d4e7451e-2bb8-4be0-bda9-90e742c39d05/dijn_a_60764_f0002_c.jpg)
Figure 3 Morphological shapes (A) and particle sizes (B) of 5-FU-loaded BSA-LTB nanoparticles.
Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.
![Figure 3 Morphological shapes (A) and particle sizes (B) of 5-FU-loaded BSA-LTB nanoparticles.Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.](/cms/asset/03aa7ccd-ec7c-4cdf-873a-e77164300126/dijn_a_60764_f0003_b.jpg)
Figure 4 In vitro release profile of BSA-LTB nanoparticles in phosphate-buffered saline (pH 5.4 and pH 7.4 at 37°C) for 48 hours.
Note: Data are expressed as the mean ± standard deviation (n=3).
Abbreviations: BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.
![Figure 4 In vitro release profile of BSA-LTB nanoparticles in phosphate-buffered saline (pH 5.4 and pH 7.4 at 37°C) for 48 hours.Note: Data are expressed as the mean ± standard deviation (n=3).Abbreviations: BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.](/cms/asset/6ecc955f-6d20-4074-9dd7-32a71d13e3ab/dijn_a_60764_f0004_c.jpg)
Figure 5 In vitro uptake ability of NPs.
Notes: Fluorescent microscopy analysis of the uptake of FITC-labeled BSA NPs and FITC-labeled BSA-LTB NPs in SMMC-7721 cells. The scale bar is 50 μm (A). Fluorescence spectrum analysis of the uptake of FITC-labeled BSA NPs and FITC-labeled BSA-LTB NPs in SMMC-7721 cells (n=3, *P<0.05, versus the corresponding BSA NPs) (B). Fluorescence spectrum analysis of the uptake of NPs labeled by FITC incubated with SMMC-7721 cells for 6 hours in media containing different amounts of free LTB (n=3, *P<0.05, versus the corresponding NP groups in media containing 5 μg/mL of LTB) (C).
Abbreviations: BSA, bovine serum albumin; FITC, fluorescein isothiocyanate; LTB, heat-labile enterotoxin subunit B; NP, nanoparticle.
![Figure 5 In vitro uptake ability of NPs.Notes: Fluorescent microscopy analysis of the uptake of FITC-labeled BSA NPs and FITC-labeled BSA-LTB NPs in SMMC-7721 cells. The scale bar is 50 μm (A). Fluorescence spectrum analysis of the uptake of FITC-labeled BSA NPs and FITC-labeled BSA-LTB NPs in SMMC-7721 cells (n=3, *P<0.05, versus the corresponding BSA NPs) (B). Fluorescence spectrum analysis of the uptake of NPs labeled by FITC incubated with SMMC-7721 cells for 6 hours in media containing different amounts of free LTB (n=3, *P<0.05, versus the corresponding NP groups in media containing 5 μg/mL of LTB) (C).Abbreviations: BSA, bovine serum albumin; FITC, fluorescein isothiocyanate; LTB, heat-labile enterotoxin subunit B; NP, nanoparticle.](/cms/asset/911fe642-e919-4797-b684-d2517a48842f/dijn_a_60764_f0005_c.jpg)
Figure 6 Viability of cells after incubation with either free 5-FU, 5-FU-loaded BSANPs, or 5-FU-loaded BSA-LTB NPs, at various 5-FU concentrations (3.75–30 μg/mL).
Notes: (A) After 24 hours; (B) after 48 hours; (C) after 72 hours (n=3)
Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B; NPs, nanoparticles.
![Figure 6 Viability of cells after incubation with either free 5-FU, 5-FU-loaded BSANPs, or 5-FU-loaded BSA-LTB NPs, at various 5-FU concentrations (3.75–30 μg/mL).Notes: (A) After 24 hours; (B) after 48 hours; (C) after 72 hours (n=3)Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B; NPs, nanoparticles.](/cms/asset/ffd380cc-0296-473c-b791-05748caf78fd/dijn_a_60764_f0006_c.jpg)
Figure 7 Cell apoptosis determined by Annexin V-FITC staining.
Notes: Flow cytometer analysis of the apoptotic and necrotic cells (Q1: necrotic; Q2: late apoptotic; Q3: live; Q4: early apoptotic) after 48 hours of incubation with the free 5-FU, 5-FU-loaded BSA NPs, and 5-FU-loaded BSA-LTB NPs, respectively. Results are expressed as means ± standard deviation (n=3).
Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; FITC, fluorescein isothiocyanate; LTB, heat-labile enterotoxin subunit B; NPs, nanoparticles; PI, propidium iodide.
![Figure 7 Cell apoptosis determined by Annexin V-FITC staining.Notes: Flow cytometer analysis of the apoptotic and necrotic cells (Q1: necrotic; Q2: late apoptotic; Q3: live; Q4: early apoptotic) after 48 hours of incubation with the free 5-FU, 5-FU-loaded BSA NPs, and 5-FU-loaded BSA-LTB NPs, respectively. Results are expressed as means ± standard deviation (n=3).Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; FITC, fluorescein isothiocyanate; LTB, heat-labile enterotoxin subunit B; NPs, nanoparticles; PI, propidium iodide.](/cms/asset/2da06704-ca6d-405b-8e2c-19f97fd419a3/dijn_a_60764_f0007_c.jpg)