Fig. 6.

Blocking csGRP78 induces lysosomal degradation of BACE2. a Co-immunofluorescence staining of csGRP78 (red) and BACE2 (green) in GSC20 and 267. Scale bar, 25 μm. b Colocalization analysis for co-expression of BACE2 and csGRP78 in cell membrane of GSC20 and 267 using colocalization finder plugin. c Western blotting for BACE2 in GSC267 with anti-GRP78 treatment for 72 h or lentiviral shGRP78 expressing. d BACE2 RNA level in GSC20 and 267 was detected by qRT-PCR with anti-GRP78 treatment or lentiviral shGRP78 expressing, GADPH as the reference gene. e Western blotting for BACE2 protein in GSC267 that treated with MG-132, Chloroquine (CQ) or anti-GRP78, respectively, and co-treatment of anti-GRP78 with MG-132 or CQ. f and g Upper, the confocal microscopy images for overview and splitting channel of BACE2 (red) and LAMP1 (green). The yellow triangles indicate the overlapped regions (yellow). Lower, the analysis plot for the fluorescence intensity of two channels. Red line for BACE2 and green line for LAMP1. Scale bar, 5 μm, and 25 μm for the large field of view. Error bar indicates at least three independent experiments and data are shown as mean ± SD