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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. 2 — PanCa-related NGF activates SC autophagy. A. NGF secretion measured by ELISAs in the serum-free medium of five pancreatic cancer cell lines (* p < 0.05). B. NGF secretion measured by ELISAs in the serum-free medium of the control and shNGF PANC-1 cancer cell lines (* p < 0.05). C. NGF secretion measured by ELISAs in the serum-free medium of the control and shNGF BxPC-3 cancer cell lines (* p < 0.05). D. LC3 immunofluorescence (green) of RSC96 cells treated with the control or shNGF-PanCa CM or PanCa-conditioned medium. White arrows: positive LC3 puncta indicating autophagic vacuoles. E. Statistics of LC3 puncta number in RSC96 cells treated with the control or shNGF-PANC-1 CM or normal PANC-1 conditioned medium (* p < 0.05). F. Statistics of LC3 puncta number in RSC96 cells treated with the control or shNGF-BxPC-3 CM or normal BxPC-3 conditioned medium (* p < 0.05). G. P62 immunofluorescence (green) of RSC96 cells treated with the control or shNGF-PanCa CM or PanCa-conditioned medium. White arrows:positive P62 punctas indicating autophagic vacuoles. H. Statistics of P62 puncta number in RSC96 cells treated with the control or shNGF-PANC-1 CM or normal PANC-1-conditioned medium (* p < 0.05). I. Statistics of P62 puncta number in RSC96 cells treated with the control or shNGF-BxPC-3 CM or normal BxPC-3-conditioned medium (* p < 0.05). J. Autophagic flux detection of RSC96 cells treated with the control or shNGF-PANC-1 CM or PANC-1-conditioned medium. LC3 labeled with GFP (green) and RFP (red) and merged as yellow. Red arrow: autophagolysosomes. Yellow arrow: autophagosome. PANC-1-conditioned medium could promote autophagy flux in RSC96 cell line while inhibiting NGF secrection in PANC-1 could partially reverse the effect. K. Autophagic flux detection of RSC96 cells treated with the control or shNGF-BxPC-3 CM or BxPC-3-conditioned medium. LC3 labeled with GFP (green) and RFP (red) and merged as yellow. Red arrow: autophagolysosomes. Yellow arrow: autophagosome. PANC-1-conditioned medium could promote autophagy flux in RSC96 cell line while inhibiting NGF secrection in BxPC-3 could partially reverse the effect. L. Statistics of red and yellow puncta number in RSC96 cells treated with the control or shNGF-PANC-1 CM or PANC-1-conditioned medium (* p < 0.05). M. Statistics of red and yellow puncta number in RSC96 cells treated with the control or shNGF-BxPC-3 CM or BxPC-3-conditioned medium (* p < 0.05). N. Western blotting of NGF in the control or NGF knockdown PANC-1 cell lines. Actin was used as a loading control. Several autophagy related proteins were detected. PANC-1-conditioned medium could inhibit p-mTOR while promote p-ULK1 and ATG5 expression in RSC96 cell line. Moreover, PANC-1-conditioned medium could promote the conversion of LC3I to LC3II and the degradation of P62. Inhibiting NGF secretion of PANC-1 by knocking down NGF could partially reverse the effect. O. Western blotting of NGF in the control or NGF knockdown BxPC-3 cell lines. Actin was used as a loading control. Several autophagy related proteins were detected. BxPC-3-conditioned medium could inhibit p-mTOR while promote p-ULK1 and ATG5 expression in RSC96 cell line. Moreover, BxPC-3-conditioned medium could promote the conversion of LC3I to LC3II and the degradation of P62. Inhibiting NGF secretion of BxPC-3 by knocking down NGF could partially reverse the effect