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. Author manuscript; available in PMC: 2023 Aug 1.
Published in final edited form as: Adv Ther (Weinh). 2022 Jun 17;5(8):2200018. doi: 10.1002/adtp.202200018

Figure 3. Cellular uptake of AuNS loaded with miRs and TK-p53-NTR plasmid DNA in 4T1 TNBC cells.

Figure 3.

(a) Confocal laser microscopic images showing the internalization of pAuNS coloaded with Cy5-tagged miR and TK-p53-NTR plasmid DNA in 4T1 cells (Red: Cy5-miR, CytoCy5S, and Blue: Hoechst 33342 for nuclear staining); (b) Fluorescent microscopic images indicating the Cy5-signal for miRs delivery and CytoCy5S substrate conversion for TK-p53-NTR expression in 4T1 cells transfected with pAuNS co-loaded with Cy5-miR and TK-p53-NTR plasmid. (c) The same results are shown in ‘b’ measured by flow cytometry analysis. (d-f) Quantitative evaluation of miRs and TK-p53-NTR plasmid transfection in 4T1 cells by AuNS for the intracellular miRs using qRT-PCR, and TK-p53-NTR by using qPCR (n =3; Two-way ANOVA with Tukey T test and confidence interval significance was performed for comparisons; * represents p < 0.05, ** represents p < 0.01, and ***represents p < 0.001; the data were plotted as mean ± SD). (g) Immunoblot analysis of the downstream target proteins of antimiR-21, antimiR-10b, and miR-100 in 4T1 cells transfected with AuNS co-loaded with miRs and TK-p53-NTR plasmid (G1: pAuNS-pcDNA; G2: pAuNS@TK-p53-NTR; G3: pAuNS@TK-p53-NTR-miRs; G4: non-targeted AuNS@pTK-p53-NTR-miRs). All the non-targeted AuNS NPs used in the experiments were conjugated with scrambled-uPA peptide (sc-uPA).