Fig. 6: JAK/STAT pathway activation by ectopic mahe induces hid dependent apoptosis. | Cell Death & Disease

Fig. 6: JAK/STAT pathway activation by ectopic mahe induces hid dependent apoptosis.

From: Maheshvara regulates JAK/STAT signaling by interacting and stabilizing hopscotch transcripts which leads to apoptosis in Drosophila melanogaster

Fig. 6

ad Activation of JAK/STAT pathway was detected by 10X STAT92E-GFP reporter in eye-antennal discs. a, c Significant increase in GFP level was observed in GMR-GAL4 driven mahe eye antennal disc when compared to the reporter line alone, indicating activation of JAK/STAT signaling (compare area in rectangle). b, d DAPI was used to mark the nucleus. el Gain of function clone of mahe displays enhanced Stat92E activity. Gain of function clones of mahe using UAS-HA-mahe flies were generated with FLP/FRT system in salivary gland. f, j GFP positive and non-GFP cells mark mahe gain-of-function clones (marked with arrow) and wild-type cells (marked with arrowhead), respectively. e Level of Stat92E the transcription factor for JAK/STAT signaling was enhanced in mahe gain-of function somatic clones when compared to wild-type neighboring cells. i Clonal area marked with GFP positive cells, shows elevated level of Mahe in the gain-of-function clones. g, k DAPI was used to mark the nucleus. h, l Fourth column represents merged image with DAPI staining. m Graph represents Stat92E intensity per unit area which shows marked enhancement in the level of Stat92E in nucleus and cytoplasm, in mahe overexpressing clones when compared to neighboring cells. One-way ANOVA. n, n Ectopic expression of mahe results in rough eye phenotype. o, o and p, p mahe induced rough eye phenotype was rescued by H99 and hid mutant, but not with rpr and grim mutants (q, q and r, r). s Real time PCR showed a threefold increase in hid transcript levels upon ectopic expression of mahe, while downregulation of JAK/STAT pathway by stat92E-RNAi rescued the hid levels. One-way ANOVA. Scale bar in 50 µm (ad, el). Genotypes (a, b) w/+; 10X STAT-GFP/+, GMR-GAL4/+ (c, d) w/+; 10X STAT-GFP/+; GMR-GAL4, UAS-HA-mahe/+ (n) w/+; +; GMR-GAL4,UAS-HA-mahe/+ (o) w/+; +; GMR-GAL4,UAS-HA-mahe/H99 (p) w/+; +; GMR-GAL4,UAS-HA-mahe/hid05014 (q) w/+; +; GMR-GAL4,UAS-HA-mahe/rpr87 (r) w/+; +; GMR-GAL4,UAS-HA-mahe/grimC15E. One-way ANOVA * P < 0.05, ** P < 0.01, *** P < 0.001.

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