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. 2021 Dec 24;19:447. doi: 10.1186/s12951-021-01189-5

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© The Author(s) 2021

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — TH-NPs demonstrated targeted inhibition of the protective autophagy and significantly enhanced chemotherapy in vitro. A GFP-RFP-LC3 dual reporter assay to analyze the blockade of autophagy flux. B The abundance of RFP-LC3 puncta was significantly enhanced over GFP puncta in OXA-treated cells, illustrating OXA triggered protective autophagy indicated by the accumulation of autolysosomes. HCQ can inhibit the autophagy flux or autophagosome–lysosome fusion (Free OXA = 20 μM, HCQ = 10 μM, the equivalent concentration of OXA or HCQ in NPs or TH-NPs). Scale bar: 10 μm. C Quantification analysis of the autophagy flux of HCCLM3 after the indicated treatments. Eight random fields (n = 8) were used for quantificationally analyzing the intracellular level of autophagosomes (GFP and RFP puncta) and autolysosomes (RFP-only puncta). D Representative TEM images of intracellular level of autophagic vacuoles in HCCLM3 cells (N, nucleus; red arrow, autolysosomes; green arrow, autophagosomes). E Quantification of the proliferation index of HCCLM3, HepG2 and Huh-7 cells according to the crystal violet staining after the indicted treatments. Data were displayed as the mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001