Fig. 6.

Eiger/TNF exocytosis activates JAK-STAT signaling through JNK. (A) The JAK-STAT activity GFP-reporter line shows pathway activation in Ras^V12 clones. The lower panel shows STAT-GFP alone. (B) dTAK1-rescued Ras^V12 clones (red) induced in dtak1 hemizygous mutant discs do not show JAK-STAT GFP (green) induction. Lower panel shows STAT-GFP alone. The red outlines show clone boundaries in A and B. (C,D) Ras^V12 or Ras^V12, dom-RNAi double mutant clones stained with DAPI to mark nuclei and with antibody against the mitotic marker, phospho-histone-3 (PH3). The majority of Ras^V12 clones are phospho-histone-3 positive (C). The lower panel of C represents a magnified view of DAPI and phospho-histone-3 from the boxed area in the upper panel. Ras^V12, dom-RNAi clones (D) show reduced phospho-histone-3 staining. The lower panel shows the phospho-histone-3 channel alone. The green outlines show clone boundaries in D. Scale bars: 5 μm. (E) Quantification of C and D. (F) RT-PCR experiment measuring Socs36E levels relative to expression levels of the housekeeping gene Rp49. Normalized fold differences in Socs36E expression levels between Ras^V12 and wild-type (WT) or between Ras^V12, eiger-RNAi (eiger RNAi-knockdown specifically in RasV12 clones) and Ras^V12 or between Ras^V12, sec15 double mutant and Ras^V12 clones are shown. Experiments were performed in triplicate and standard deviations were derived from the coefficient variations of experimental and control samples.