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. 2021 Feb 1;4(3):e202000693. doi: 10.26508/lsa.202000693

Figure 4. Glioma-secreted ImpL2 inhibits insulin pathway activity in neurons.

Figure 4.

(A, B, C) Confocal microscopy images of adult brain of 7-d-old flies from (A) repo>UAS-LacZ (Control), (B) repo>UAS-dEGFRλ, UAS-dp110CAAX (Glioma) and (C) repo>UAS-dEGFRλ, UAS-dp110CAAX, UAS-ImpL2 RNAi (Glioma ImpL2↓) genotypes, combined with tGPH transgene reporter of PI3K activity (green). (A′, B′, C′) Merge with glial nuclei in red and DAPI staining (scale bar 10 μm). White arrows point to non-glial cells with PI3K activity. The images are representative of at list N = 6 per genotype. (D, E, F) Confocal microscopy images of 7-d-old adult fly brains of the genotypes: (D) repo>UAS-LacZ, (E) repo>UAS-dEGFRλ, UAS-dp110CAAX and (F) repo>UAS-dEGFRλ, UAS-dp110CAAX, UAS-ImpL2 RNAi, in all the cases combined with ThorMI09732 line. (D′, E′, F′) Neuronal nuclei are marked in blue. White arrows indicate nuclear localization of ThorGFP signal. (G) Quantification and statistical analysis of co-localization area in N = 9 per genotype (ANOVA, post hoc Bonferroni) (scale bar, 50/10 μm). (H, I, J, K) Adult neuromuscular junction (NMJ) from the genotypes: (H) D42>UAS-LacZ (Control), (I) D42>UAS-InRDN (Neuron InRDN), and (J) repo>UAS-ImpL2 (Glia ImpL2↑) animals after 7 d at 29°C (active zones shown in green). D42 is expressed in motor neurons. (K) Quantification and statistical analysis of active zones in at least N = 6 per genotype (ANOVA, post hoc Bonferroni) (scale bar, 50 μm) (***P-value < 0.001).