Fig. 2.
Activation of JNK signaling in Nact/lgl-IR tumor. Fluorescent micrographs of wing imaginal discs are shown. a Endogenous expression of puc in Drosophila wing imaginal disc is shown. b Only Nact overexpression shows slight increment in puc expression. c Downregulation of lgl in wing disc induces puc expression. d A significant increase in puc expression was found, when Nact is coexpressed with lgl-IR. a′–d′ are the merge images of DAPI along with a–d, respectively. Images (e–h) show the expression of Egr in wild-type, Nact, lgl-IR, and Nact/lgl-IR wing discs, respectively. The expression of Egr remains unchanged in all genotypes. e′–h′ are the merge images of DAPI along with e–h. i Analyses of differences in wing disc size show that there is a significant increase in Nact/lgl-IR wing imaginal disc size as compared to the wild-type, Nact, and lgl-IR wing disc sizes. j Real-time PCR analysis was performed to estimate the transcript levels of JNK pathway ligand egr and its receptors wgn and grnd in Nact/lgl-IR tumor. The transcript levels of egr and wgn in Nact/lgl-IR were significantly reduced, when compared to that of wild-type; however no significant change in transcript levels were seen, when compared to that of only Nact and lgl-IR. Interestingly, grnd transcript levels were significantly upregulated in Nact/lgl-IR tumor as compared to that of the wild-type, Nact and lgl-IR wing discs. qPCR was normalized with rps17 and repeated for three times. Analysis of data was done using two-way ANOVA with Tukey’s multiple comparison test; data represents mean ± SEM (***p < 0.001; **p < 0.01; *p < 0.05 and ns p > 0.05). All wing discs are oriented with dorsal to the top and posterior to the right. Scale bars: 100 µm (a–d, a′–d′, e–h, e′–h′)