Fig. 5. JNK mediates IRE1-induced autophagy-dependent neuron death.

a Immunoblot analysis of the phosphorylation levels of JNK from the head lysates of adult GMR-Gal4 > + versus GMR-Gal4 > IRE1 flies (n = 30 flies/genotype; representative of three independent experiments). Scale bar represents 30 µm. b Confocal microscopy analysis of eye discs from GMR-Gal4 > puc^E69 versus GMR-Gal4 > puc^E69; IRE1 larvae. Shown are representative images for the JNK reporter puc-lacZ expression by immunostaining with anti-β-Gal antibody, along with DAPI staining (n = 20 flies/genotype). The enlarged regions are indicated. Scale bar represents 30 µm. c Immunoblot analysis of JNK phosphorylation in the adult head lysates of GMR-Gal4 > +, GMR-Gal4 > IRE1 and GMR-Gal4 > IRE1; Bsk-Ri lines (n = 30 flies/genotype; representative of three independent experiments). (d) Representative light microscopy (left panels) and SEM (middle and right panels) images of external eyes from adult GMR-Gal4 > IRE1 and GMR-Gal4 > IRE1; Bsk-Ri versus GMR-Gal4 > + and GMR-Gal4 > Bsk-Ri lines (n = 5–8 flies/genotype). Scale bar represents 50 µm. e Cell death analysis of larval eye discs from the indicated lines. Shown are representative images of TUNEL and DAPI staining along with IRE1 immunostaining (n = 20 flies/genotype). Scale bar represents 100 µm