Fig. 1: Muscleblind (Mbl) is a novel modifier of FUS toxicity in vivo. | Nature Communications

Fig. 1: Muscleblind (Mbl) is a novel modifier of FUS toxicity in vivo.

From: Muscleblind acts as a modifier of FUS toxicity by modulating stress granule dynamics and SMN localization

Fig. 1: Muscleblind (Mbl) is a novel modifier of FUS toxicity in vivo.The alternative text for this image may have been generated using AI.

An unbiased genetic screen using our Drosophila model of ALS revealed candidate modifiers of toxicity caused by expression FUS. a Schematic of the Df(2 R)Exel6066 line identified in the screen and the relative locations of the genes within the deficient region. The Df(2 R)BSC154 deficiency region overlapped with mbl, beginning in the middle of the gene. b Representative panel of adult Drosophila eyes showing degeneration caused by expression of wild-type and ALS-linked mutant FUS with and without concurrent heterozygous depletion of the genes within the indicated deficiency lines. c Quantification of eye degeneration severity in Drosophila indicating significant suppression of FUS-associated toxicity when these flies are crossed with the deficiency lines (****P < 0.0001, *P = 0.0160). d Representative images of Drosophila eyes expressing the indicated FUS proteins alone (left column), in combination with RNAi-mediated knockdown of mbl (middle column), or with overexpression of Mbl isoform C (right column). (N = 17–117). e Quantification of eye degeneration severity confirms significant suppression of FUS toxicity following depletion of mbl (left graph), and significant enhancement of toxicity following overexpression of MblC (right graph) (****P = 0.0001, ***P = 0.0003). f qPCR of RNA (n = 3) confirms significant knockdown of endogenous mbl in the RNAi line (**P = 0.0021). g Western blot (WB) analysis following co-immunoprecipitation (Co-IP) experiments using HEK293T cells. Co-IP was performed using anti-FUS antibody to target endogenous FUS (n = 3). WBs were probed for human MBNL1, which was present in the input control and the immunoprecipitated samples but absent from the negative control containing only beads. h WB showing FUS and tubulin protein levels in Drosophila expressing wild-type and mutant FUS (N = 3). i Quantification of FUS normalized to tubulin, indicating equivalent FUS expression in all FUS-expressing groups. In addition, knockdown of Mbl has no effect on FUS levels (NS = not significant, P = 0.3578). Statistical significances in c and e were determined using two-tailed t tests for each FUS pair (Mann–Whitney test). One-tailed t test was used in f. One-way ANOVA with Tukey’s multiple comparisons test was used for h. All quantifications are represented as the mean ± SD.

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