Fig. 10 | Scientific Reports

Fig. 10

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. 10

(a) Fluorescence microscopy images of untreated control cells, cells treated with MTX, cells treated with MTX-SLNs, and cells treated with MTX-Lf-SLNs displaying Caspase-6 (green, Alexa Fluor 488) and nuclei (blue, DAPI). Untreated control cells exhibited minimal Caspase-6 expression, leading to low green fluorescence (12.4 ± 1.3 RFI). MTX therapy leads to strong green fluorescence (45.6 ± 3.2 RFI, p < 0.001), suggesting significant apoptosis and nuclear breakdown. Decreased fluorescence (32.8 ± 2.6 RFI, p < 0.01 compared to MTX) in cells treated with MTX-SLNs indicates that controlled drug release reduces direct cytotoxic effects. Cells exposed to MTX-Lf-SLNs displayed diminished green fluorescence (20.2 ± 1.8 RFI, p < 0.001 compared to MTX; not significant when compared to control), indicating receptor-mediated targeting and minimal apoptosis. (b) Quantitative analysis of fluorescence intensity for Caspase-6 (green, Alexa Fluor 488) and nuclei (blue, DAPI). Findings show considerable differences in Caspase-6 expression among treatments (ANOVA, p < 0.001). DAPI fluorescence stayed stable, showing consistent cell density. The combined fluorescence intensities reflected Caspase-6 activity, providing additional support for the link between apoptosis and type of treatment.

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