Figure 2.
Hsp83 and Cdc37 Are Required for the Activation of InR Pathway
(A) Larval brains of control (UAS-dicer2), hsp83 RNAi (v108568), and cdc37 RNAi (v47776) under insc-Gal4 at 72 hr ALH were stained for Mira, Dpn, and Akt. Yellow dotted boxes indicate the region of zoomed-in images. Central brain is to the left of white dotted lines. White arrows point to NSCs and outlines of NSCs are indicated by white dotted lines. Scale bars, 30 μm for whole brain lobe and 5 μm for single cell.
(B and C) Western blot analysis of larval brain extracts of control (UAS-dicer2), hsp83 knockdown (v108568), and cdc37 knockdown (v110727) induced with insc-Gal4 at 72 hr ALH. Blots were probed with anti-Hsp83 antibody or anti-Akt antibody.
(D and E) Western blot analysis of larval brains of control (UAS-dicer2), hsp83 knockdown (v108568), cdc37 knockdown (v110727), and InRAD overexpression (O/E; induced with insc-Gal4) at 72 hr ALH. Blots were probed with anti-pInR antibody.
(F and G) Western blot analysis of whole larvae extracts of control (UAS-dicer2), hsp83-HA overexpression induced with insc-Gal4 at 6 hr ALH. Blots were probed with anti-Akt antibody.
(H and I) Western blot analysis of whole larvae extracts of control (UAS-dicer2), hsp83-HA overexpression induced with insc-Gal4 at 6 hr ALH. Blots were probed with anti-pInR antibody. Loading control, actin (B, D, F, and H).
Statistical analyses were done comparing between two different genotypes using a two-tailed Student's t test (C, E, G, and I). ∗p < 0.05, ∗∗∗p < 0.001. Data are presented as mean ± SD in (C), (E), (G), and (I).