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. 2025 Jul 11;15(16):7973–7989. doi: 10.7150/thno.112649

Figure 2.

Figure 2

In vitro evaluation of Nrf2 silencing, proliferation inhibition, and apoptosis induction by NPs(3-MA/siNrf2/P-18). (A, B) Quantitative real-time PCR (qRT-PCR) analysis of relative Nrf2 mRNA expression in MDA-MB-231 and 4T1 cells, respectively, following treatment with NPs(3-MA/siNrf2/P-18). (C, D) Western blot analysis of Nrf2 protein levels in MDA-MB-231 and 4T1 cells after the indicated treatments. (E, F) Cell proliferation assays showing growth inhibition in MDA-MB-231 and 4T1 cells treated with NPs(3-MA/siNrf2/P-18) with or without ultrasound (US) irradiation. (G) Representative colony formation assay images and (H, I) Quantitative analysis of colony numbers in MDA-MB-231 and 4T1 cells, respectively, following treatment with NPs(3-MA/siNrf2/P-18) under US irradiation. (J-M) Flow cytometry analysis of apoptosis rates in MDA-MB-231 and 4T1 cells after treatment with NPs(3-MA/siNrf2/P-18) with US activation. G1: Blank; G2: NPs(3-MA/siNrf2/P-18); G3: NPs(3-MA/siCTL/P-18) + US; G4: NPs(G0-C14/siNrf2/P-18) + US; G5: NPS(3-MA/siNrf2/P-18) + US. Data are presented as mean ± SD (n = 3). Error bars represent standard deviation. Statistical analysis was performed using one-way ANOVA for multiple comparisons. Significance is indicated as *P < 0.05, **P < 0.01 and ***P < 0.001, ns, no significance.