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. 2019 Dec 17;17(12):e3000545. doi: 10.1371/journal.pbio.3000545

Fig 7. JNK pathway is necessary to localize Fz1 in the TMs, grnd mediates Wg pathway activation in GB and promote GB progression.

Fig 7

Brains from third instar larvae displayed at the same scale. Glia are labeled with UAS-Ihog-RFP (gray or red in the merge) driven by repo-Gal4 to visualize active cytonemes/TM structures in glial cells and stained with Fz1 (gray or green in the merge, panels A–F) and Cyt-Arm (gray or green in the merge, panels G–K) in the following genotypes control, glioma, glioma grnd-RNAi, Glioma grnd−/− and Glioma BskDN brain sections. Fz1 and Cyt-Arm average pixel intensity staining quantification ratio between ihog+ and ihog–domains (F and L). (A, G) Fz1 and Cyt-Arm is homogeneously distributed in control sections. (B, H) In glioma brains, Fz1 accumulates in the TMs, and Cyt-Arm accumulates in the neurons’ cytoplasm where it is inactive. (C–D, I–J) Knockdown or knockout of grnd and BskDN in glioma brains showing a normal glial network, and Fz1 and Cyt-Arm does not accumulate showing a homogeneous distribution similar to the control. Fz1 average pixel intensity quantification for glia/neuron ratio (F) and Cyt-Arm average pixel intensity quantification for neuron/glia ratio between ihog- and ihog+ domains (L). Nuclei are marked with DAPI (blue). Error bars show SD; *P < 0.01, **P < 0.001, ***P < 0.0001, or ns for nonsignificant. Scale bar size is indicated in this and all figures. The data underlying this figure can be found in S1 Data. Genotypes: (A, G) repo-Gal4, ihog-RFP/UAS-lacZ, (B, H) UAS-dEGFRλ, UAS-dp110CAAX;; repo-Gal4, UAS-ihog-RFP, (C, I) UAS-dEGFRλ, UAS-dp110CAAX; UAS-grnd-RNAi; repo-Gal4, UAS-ihog-RFP, (D, J) UAS-dEGFRλ, UAS-dp110CAAX; grndMINOS/grndMINOS; repo-Gal4, UAS-ihog-RFP, (E, K) UAS-dEGFRλ, UAS-dp110CAAX;; repo-Gal4, UAS-ihog-RFP/ UAS-bskDN. BskDN, dominant negative form of the effector Bsk; Cyt-Arm, Cytoplasmic-Armadillo; Fz1, Frizzled1; GB, glioblastoma; grnd, Grindelwalkd; Ihog, interference hedgehog; JNK, cJun N-terminal kinase; RFP, Red Fluorescent Protein; TM, tumor microtube; Wg, wingless.