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