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. 2022 Jun 6;12(10):4791–4801. doi: 10.7150/thno.69168

Figure 3.

Figure 3

Cellular uptake and cytotoxicity of DOX and DOX prodrugs. (A) Fluorescence microscopy images of 4T1 breast cancer cells incubated with 25 µM of DOX, DOX-AU1, and DOX-AU2 for 4 h in the presence and absence of β-GUS (scale bar 100 µM). (B-C) Quantification of intracellular fluorescence for DOX and DOX prodrugs in the presence and absence of β-GUS compared to untreated cells (Ctrl). (D-G) Viability of 4T1 breast cancer cells treated with DOX and DOX prodrugs pulse-incubated for 4 h (followed by 68 h incubation in medium; D, F) and incubated continuously for 72 h (E, G), in the presence (F, G) and absence (D, E) of β-GUS. Statistical differences were determined using a one- way (B, C) and two-way ANOVA with multiple comparison (D-G). *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.