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Conjugation of MTX to ND prolongs MTX retention in breast cancer cells. (A) Representative images of MTX retention at (i) 6 h and (ii) 24 h after 1 h of treatment with IC50 concentrations of MTX and ND–MTX. ND–MTX treatment at 24 h showed significant MTX retention (indicated by red arrows). (B) Quantitative scores of MTX retention efficiency at (i) 6 h and (ii) 24 h after 1 h of treatment with IC50 concentrations of MTX and ND–MTX.**, p < 0.01; *, p < 0.05. Scale bar represents 50 μm.
