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. 2020 Dec 10;9:e59419. doi: 10.7554/eLife.59419

Figure 5. Modulation of nucleocytoplasmic transport rescues autophagolysosome dysfunction.

(A) Drosophila larval salivary glands stained with anti-Mitf and DAPI expressing +/- UAS-30R, UAS-shuttle-GFP (S-GFP, not shown), and either control RNAi (UAS-lucRNAi) or exportin RNAi (UAS-embRNAi ) under the control of vGlut-Gal4. Scale bar = 10 µm (B) Quantification of percent (%) nuclear Mitf in A. Data are reported as mean ± SEM. Student’s t-test, n = 4 and 5 larvae, respectively. (C) Drosophila motor neurons expressing UAS-GFP:Lamp1 (N-terminal, luminal GFP) -/+ UAS-30R and UAS-lucRNAi or exportin RNAi (UAS-embRNAi ). Scale bar = 10 µm. (D) Quantification of C. Student’s t-test, n = 6 larvae. (E) Drosophila motor neurons expressing UAS-mCherry:Atg8 +/- UAS-30R and either control RNAi (UAS-lucRNAi) or exportin RNAi (UAS-embRNAi ). Scale bar = 10 µm. (F) Quantification of E. Data are reported as mean ± SEM. Mann-Whitney test, n = 10 larvae. (G) Drosophila motor neurons expressing UAS-p62:GFP -/+ UAS-30R and either control RNAi (lucRNAi) or exportin RNAi (embRNAi ) under the control of vGlut-Gal4. Scale bar = 10 µm. (H) Quantification of G. Data are reported as mean ± SEM. Brown-Forsythe and Welch ANOVA test, p<0.0001, followed by Dunnett’s T3 multiple comparisons, n = 12–14 larvae per genotype.

Figure 5.

Figure 5—figure supplement 1. Nucleocytoplasmic transport disruption is upstream of autophagic defects.

Figure 5—figure supplement 1.

(A) Images of the nucleocytoplasmic transport marker shuttle-GFP (S-GFP) in control and vGlut > 30R with either UAS-luciferase (control) or UAS-Ref(2)P RNAi #1 expressed in Drosophila salivary glands. (B) Images of L3 salivary gland expressing UAS-S-GFP in vGlut > 30R Drosophila after feeding supplemented with control (DMSO), 0.2% trehalose, or 1.0 µm rapamycin. (C) Representative images of ventral nerve cords expressing control, knockdown of RanGAP (UAS-RanGAPRNAi), or overexpression of RanGAP (UAS-RanGAP), along with UAS-p62:GFP in vGlut motor neurons. Scale bars = 10 µm.