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. 2021 Jan 7;40:20. doi: 10.1186/s13046-020-01807-4

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© The Author(s) 2020

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 — csGRP78 is preferentially expresses in MES GSCs and regulates stemness of GSCs. a csGRP78 was detected by nonpermeabilized immunofluorescence in GSC11, 8–11, 20 and 267. Scale bar, 25 μm. b csGRP78 in GSCs were quantified by flowcytometry, and the mean fluorescence intensity of PE (FL2 channel) represents the expression of csGRP78. c Western blotting for total GRP78 and MES-related signatures and pathways after GSC20 and 267 treated with anti-GRP78. d Upper, proportion of apoptotic cells and lower, G2/M phase cells of GSC20 and 267 treated with anti-GRP78. e Images of tumor spheres formation after blocking csGRP78 in GSC20 and 267 and quantified with diameter of spheres. Scale bar, 500 μm. f In vitro limiting dilution assay for GSC20 and 267 treated with anti-GRP78 g Left, nonpermeabilized immunofluorescence of csGRP78 in NPC and right, csGRP78 expression detected by flowcytometry. Scale bar, 25 μm. h Apoptosis of NPCs after anti-GRP78 treatment was detected by flowcytometry. i Western blotting for phospho-STAT3 and STAT3 protein expression in NPCs after anti-GRP78 treatment. Error bar indicates at least three independent experiments and data are shown as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001