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. 2022 Feb 2;41:48. doi: 10.1186/s13046-021-02198-w

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© The Author(s) 2022

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. 5 — The effect of double targeting of NGF and autophagy on cancer cells. A. CCK-8 proliferation assay of PANC-1 and BxPC-3 cells treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM). The OD 450 nm value was determined at 0 h, 24 h, 48 h and 72 h (* p < 0.05). B. Transwell migration assay of PANC-1 and BxPC-3 cells treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) at 24 h. PANC-1 or BxPC-3 were seeded in the upper chamber with 1% serum and the indicated drugs. The lower chamber were 10% serum culture medium. Black arrow: migrated cell. C. Statistics of migrated BxPC-3 cells treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. 5B) (* p < 0.05). D. Statistics of migrated PANC-1 cells treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. 5B) (* p < 0.05). E. Statistics of LC3 puncta number in PANC-1 cells treated with control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. S5A) (* p < 0.05). F. Statistics of the P62 puncta number in PANC-1 cells treated with control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. S5B) (* p < 0.05). G. Statistics of LC3 puncta number in BxPC-3 cells treated with control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. S5C) (* p < 0.05). H. Statistics of P62 puncta number in BxPC-3 cells treated with control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. S5D) (* p < 0.05). I. Statistics of cleaved PARP1-positive and cleaved PANC-1-negative cell numbers treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. S5E) (* p < 0.05). J. Statistics of cleaved caspase-3-positive and cleaved caspase-3-negative PANC-1 cell numbers treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. S5F) (* p < 0.05). K. Statistics of cleaved PARP1-positive and cleaved PARP1-negative BxPC-3 cells treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. S5G) (* p < 0.05). L. Statistics of cleaved caspase-3-positive and cleaved caspase-3-negative BxPC-3 cell numbers treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) (cell image: Fig. S5H) (* p < 0.05). M. TEM image of PANC-1 and BxPC-3 cells treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) at 24 h. Autophagy and apoptosis signs were detected. Green arrow: autophagosome. Red arrow: apoptosis sign. Yellow arrow: dysfunctional lysosome. N. Western blotting of PANC-1 and BxPC-3 cells treated with the control, RO (1 μM), CQ (40 μM) or RO + CQ (1 μM + 40 μM) at 24 h. GAPDH was used as a loading control