Fig. 6

The effects of early repression of AGR2 in both Jed66_GB and Jed41_GB. A i) Images showing minor repression of AGR2 in both cell lines following the short post-transfection of eight hours for Jed66_GB and 36 h for Jed41_GB. ii) The AGR2 mean intensities for negative control and AGR2 exon 2B siRNA-transfected cells. The asterisk indicates significant T-test difference at p < 0.05. B GRP78 expression and AGR2 expression in both negative control and AGR2 exon 2B siRNA-transfected cells for both cell lines. C Immunofluorescence images of nuclear Caspase-3 in negative control and AGR2 exon 2B siRNA-transfected cells. The arrow points to a Caspase-3-positive multinucleated cell. Caspase-3 is shown in red and DAPI in blue. D i) Images show multinucleation in Jed66_GB ii) The percentage of multinucleated cells in Jed66_GB following negative control and AGR2 exon 2B siRNA transfections. Asterisk indicate T-test significant difference at p < 0.05. E i) Immunofluorescence images displaying the expression of P53 in both cell lines following negative control and AGR2 exon 2B siRNA transfections. P53 is shown in green and DAPI in blue. ii) Magnified images showing P53 localisation. iii) The percentage of cells positive for nuclear P53 was significantly higher in Jed66_GB compared with Jed41_GB. Three asterisks indicate T-test significant difference at p < 0.001. F A suggested model that explains the role of AGR2 in glioblastoma. Both in situ and in vitro, AGR2 and GRP78 are highly expressed in CSCs and drug-resistant cells. Upon repression of AGR2, GRP78 was also reduced. Early cell fate seems to be dependent on the used model, and can lead either to multinucleation followed by cell death or to delayed growth followed by attenuated cell death