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Fig. 4 | Journal of Nanobiotechnology

Fig. 4

From: Biodegradable reduce expenditure bioreactor for augmented sonodynamic therapy via regulating tumor hypoxia and inducing pro-death autophagy

Fig. 4

Pro-death autophagy induced by HHBP promoted cancer cell apoptosis. a The relative viabilities of 4T1 cells after different treatments, quantified by CCK-8 assay (n = 6). ***P < 0.001. b Fluorescence images of 4T1 cells stained by Calcein-AM (green) and PI (red) after treatment with PBS, HMME@HMONs, HHBP without or with US irradiation. Scale bar = 200 μm. c Flow cytometry analysis of annexin V-FITC/PI-stained 4T1 cells after treatment with PBS, HMME@HMONs or HHBP without or with US irradiation. d Western blotting of the p62, LC3-I, and LC3-II levels in 4T1 cells after different treatments. (I) PBS, (II) HHBP, (III) HMME@HMONs, (IV) HHBP + US, and (V) HMME@HMONs + US. e Representative immunofluorescence staining of LC3 punctate dots and MDC-stained fluorescent images after 4T1 cells were treated with HMME@HMONs or HHBP for 4 h with or without US irradiation. Scale bars = 100 μm. f TEM images showing the formation of autophagosomes after the 4T1 cells were treated with HMME@HMONs or HHBP for 4 h with or without US irradiation. Scale bars = 2 μm. The yellow arrows in the magnification of TEM images represent the typical structures of autophagosomes. Scale bars = 1 μm

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