Fig. 7 | Scientific Reports

Fig. 7

From: Study of caspase-6 activity in aggressive HCT116 cells using methotrexate-encapsulated lactoferrin-conjugated solid lipid nanoparticles via in silico and in vitro approaches

Fig. 7

(a) Determining apoptosis via flow cytometry: Representative scatter plots and histograms illustrating the percentages of early and late apoptotic stages and changes in the cell cycle post-treatment: The sub-G1 peak significantly rose in MTX-Lf-SLNs, which exhibited the highest percentages of apoptosis and DNA fragmentation. (b) Quantitative assessment of apoptosis phases: Evaluation of the proportion of cells in various apoptotic phases (control, MTX, MTX-SLNs, MTX-Lf-SLNs). MTX-Lf-SLNs demonstrated a markedly increased rate of apoptosis (p < 0.0001) during both early and late stages. (c) DAPI fluorescence imaging of nuclear structure: the control cells exhibited intact nuclei, whereas MTX-treated cells displayed nuclear condensation and fragmentation. MTX-Lf-SLNs therapy exhibited the most pronounced chromatin condensation and nuclear shrinkage, characteristics associated with advanced apoptosis. (d) Analysis of nuclear area quantitatively with ImageJ: Treatment with MTX-Lf-SLNs resulted in a reduction of nuclear area by 18.52 ± 3.67% compared to the control, demonstrating a significantly greater apoptotic efficacy than MTX (60.34 ± 5.27%) and MTX-SLNs (42.56 ± 8.27%).

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