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. 2023 Nov 21;11:1301913. doi: 10.3389/fcell.2023.1301913

FIGURE 3.

FIGURE 3

Size compensation in the absence of Wg, JAK/STAT or JNK activity. (A) Schematic representation of the experiment used to assay the role of Wg, JAK/STAT and JNK pathways in size compensation. Larvae of the genotype indicated in the figure were kept at 17°C for 3 days after egg laying before IR. After irradiation they were kept for 4 h at 17°C to allow apoptosis and then switched to 29°C to induce the expression of the UAS transgenes that suppress the function of JNK, JAK/STAT or Wg (see Material and Methods for details). The larvae were maintained at 29°C for the rest of development until the wandering stage and then dissected and wing discs fixed. (B,B’) Confocal image of a third instar disc of the genotype indicated stained for apoptosis (Dcp1, red/white) 4 h after IR. The anterior compartment is labelled green with the Ci antibody. Note that the amount of apoptosis, monitored by Dcp1 (red/white), is the same in the anterior and posterior compartment. (C) Quantification of the levels of apoptosis 4 h after irradiation in of disc with genotype describe in (B). The percentage of Dcp1 stain is similar in A and P compartments. (D–P) Examples of confocal images of wing discs used for the quantification, GFP (green) labels the posterior compartment, and TOPRO-3 (blue) the entire disc. (D) Graph showing the volume of the posterior compartment with respect to the total volume of the disc in the different genotypes studied. There are not significant differences in posterior compartment volume between irradiated and no-irradiated larvae for all the cases. The discs in which the JAK/STAT pathway is suppressed the posterior compartments are smaller, but in both irradiated and no-irradiated discs. (E–P) Confocal images of example imaginal discs used for the quantification. Non-irradiated discs upper images, irradiated ones lower images. (E,F) control discs. (G–J); discs in which JNK pathway is supressed in the posterior compartment by the expression of (G,H) puc or (I,J) UAS-bskRNAi; (K–N) discs in which JAK/STAT pathway is supressed in the posterior compartment by (K,L) UAS-STATRNAi and (M,N) UAS-DomeRNAi; and (O,P) discs in which Wg pathway is supressed in the posterior compartment by UAS-wgRNAi,.