EMBO/SourceData huggingface dataset:
Figure 5(A) TMT-based MS results of chromatin fractions from SU-DHL-5 control and SU-DHL-5 SLF2KO cells. Volcano plot depicting depleted chromatin-associated proteins. Significant hits are shown by red dots (depleted in SLF2KO cells). Proteins belonging to the DDR and cell cycle regulation are labeled. Experiments were performed in triplicates.(E) Immunoblot analysis of human SU-DHL-5 control and SLF2KO whole cell fractions with the indicated antibodies. (F) Immunoblot analysis of human chromatin fractions of SU-DHL-5 control and SLF2KO cell lines with the indicated antibodies.
Figure 6(D) Quantification of the SAE1 and SAE2 western blots from Figure 6C. Protein expression of SAE1 and SAE2 was normalized to β-Actin protein expression. Data are presented as mean +/- SD of n=3 independent experiments. P-value determined by unpaired t-test.(E) SUMO inhibitor (SUMOi (ML-093); 0, 6.25, 12.5, 25, 50, 100, 200, 400, 800 nM) dose response curves of SLF2KO and control SU-DHL-5 cells. Cells were treated with increasing concentrations of SUMOi for 72 hours and viability was determined by negative DAPI staining and flow cytometry measurement. Error bars represent SD from three independent experiments.(F) Quantification of flow cytometry results for DAPI and Annexin V staining in human SLF2KO and control SU-DHL-5 cells after 12.5 nM of SUMOi treatment for 72 hours. P-value determined by unpaired t-test. Error bars represent SD from three independent experiments.
Figure 7(B) Immunoblot analysis of human SU-DHL-5 control and CLSPNKO cell lines upon DRB treatment (0.5µM) for indicated time points with the indicated antibodies.(C) Quantification of the p-CHK1 western blots from Figure 7B. Protein expression of p-CHK1 was normalized to β-Actin protein expression. p-CHK1 expression in control cells at 3 hours of doxorubicin treatment was arbitrarily set to 1. Data are presented as mean +/- SD of n=3 independent experiments. P-value determined by two-way ANOVA; Šídák's multiple comparisons test.(F) SUMO inhibitor (SUMOi, TAK981; 0, 6.25, 12.5, 25, 50, 100, 200, 400, 800 nM) dose-response curves of CLSPNKO and control SU-DHL-5 cells. Cells were treated with increasing concentrations of SUMOi for 72 hours and viability was determined by negative of DAPI staining and flow cytometry measurement. Error bars represent SD from three independent experiments. (G) Mafosphamide dose-response curves of CLSPNKO and control SU-DHL-5 cells. Cells were treated with mafosphamide (MAF; 0, 31.25, 62.5, 125, 250, 500, 1000, 2000, 4000 ng/ml) for 48 hours and viability was determined by negative of DAPI staining and flow cytometry measurement. Error bars represent SD from three independent experiments.
Figure 8(B) Representative histograms of the flow cytometry experiment described in Figure 8A.(G) Quantification of cell viability measured by celltiterGlo from primary murine Eµ-Myc lymphoma derived cells co-treated with indicated concentrations of SUMOi and Rabusertib for 72 hours. Bar plots represent cell viability relative to the DMSO treated control cells. Error bars represent SD from three independent experiments. P-value was determined by ANOVA test; Tukey's post hoc test.(J) Tumour size was measured over time (n=4 mice in each group). Error bars represent SEM. P-value determined by unpaired t-test.
Figure 1A. Representative bright field micrographs of netrin-1 immunohistochemistry in formalin-fixed paraffin-embedded human ovarian, endometrial and hepatocellular carcinoma (HCC) tumor sections. Scale bar 50 µm. B. Quantification by immunoblots of netrin-1 in the ECM of cancer cells of human tumors(n=3, error bars are SD). The vertical line corresponds to the intercellular zoneF. Analysis of h-netrin-1 (human recombinant netrin-1) binding to ECM components: mouse laminin-I (m-laminin-I), recombinant human vitronectin, recombinant human lumican, recombinant human integrin α6β4, recombinant human glypican-1 and recombinant human FLTR1 by bio-layer interferometry assays
Figure 2C. NP137 fragments were produced by enzymatic cleavage and subjected to electrophoresis in denaturing or non-denaturing conditions.D. Bio-layer interferometry analysis of netrin-1/NP137 and derivates after NODAGA bioconjugation. Numbers indicate the concentration in nM of NP137 and derivates.
Figure 3D. Biodistribution properties of NP137-NODAGA-111In in a Balb/cJ mouse bearing 4T1 xenografts at 48 h (4 mice), 72 h (4 mice) and 96 h (3 mice). Radioactivity incorporation was quantified by the percentage of the injected dose per gram of organ. Error bars indicate s.d..G. Whole body autoradiography of NP137-NODAGA-111In in Balb/cJ mice bearing a 4T1 (top) or an EMT6 (bottom) tumor using a beta imager system.H. Analysis of the autoradiography ratio of NP137-NODAGA-111In in tumor versus bone in Balb/cJ mice bearing EMT6 (n= 14 relative values) or 4T1 (n=24 relative values) tumors. Whiskers indicate min to max, Central band of the box indicates the median, bottom of the box indicates Q1 and top of the box indicates Q3.
Figure 5B. Detection of netrin-1 expression in the MMTV/neuT model by immunoblots (Tumor 1 to 3).E. Biodistribution properties of NP137-NODAGA-111In in MMTV/neuT mice at 96 h (3 mice) and measured for all organs. Radioactivity incorporation was quantified by the percentage of the injected dose per gram of organ. Error bars indicate s.d.F. NP137-177Lu enhances MMTV/neuT mouse survival. Mice were treated at 3.5 months with PBS, DOTA-NP137 or NP137-177Lu. Kaplan-Meier survival curves of mice treated or not with NP137 and considered as dead when cumulative tumors volume reached 2,000 mm3. Mantel Cox test; n = 10 animals/group for PBS; n = 14 animals/group for DOTA-NP137; n = 8 animals/group for NP137-177Lu; p < 0.001 between PBS and NP137-177Lu and between DOTA-NP137 and NP137-177Lu.
Figure 1(B, C) SH-SY5Y cells were infected with HSV-2 (MOI, 1), for the indicated time intervals, and culture supernatants were isolated for measurement of LDH (B) or HMBG1 (C). Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗p ≤ 0.01; ∗∗∗p≤0.001; **** p≤0.0001. (Mann-Whitney test, two-tailed in(E) SH-SY5Y cells were infected with HSV-2 (MOI=1) for the indicated time intervals, and cell lysates were immunoblotted for p-MLKL and vinculin. For comparison, we used lysates from THP1 cells treated with Z-VAD (Z, 20 μM), TNFα (T, 100 ng/ml), Birinapant (B, 100 nM), and Necrostain-1 (NEC-1, 10 μM) as indicated.(F, G) SH-SY5Y cells were infected with HSV-1 and HSV-2 (both MOI=1) for the indicated time intervals. Cell lysates were evaluated for CASP3 activity (G) and immunoblotted for cleaved caspase 3 (CL-CASP3), cleaved PARP (CL-PARP), and vinculin (F). Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗p ≤ 0.01; ∗∗∗p≤0.001; **** p≤0.0001. (Mann-Whitney test, two-tailed in(J, K) SH-SY5Y cells were infected with HSV-1 and HSV-2 (both MOI=1) for the indicated time intervals, and cell lysates were immunoblotted for GSDME and vinculin.(L, M) SH-SY5Y cells were infected with EMCV and VSV (both MOI=1) for 16 h, and cell lysates were immunoblotted for GSDME and vinculin.
Figure 2ESC-derived neuron progenitor cells were infected with HSV-1 and HSV-2 (both MOI=1) for the indicated time intervals, and lysates were isolated and analyzed as indicated by (B) immunoblottingHuman brain slices were infected with HSV-1-GFP (1˟106 PFU/ml) and HSV-2-GFP (1˟106 PFU/ml) for 72 h, visualized microscopicallyHuman brain slices were infected with HSV-1-GFP (1˟106 PFU/ml) and HSV-2-GFP (1˟106 PFU/ml) for 72 h, and lysed for analysis of GSDME and vinculin.Mouse mixed brain cells were infected with HSV-2 (MOI=10) for 36 h and lysed for analysis of GSDME and vinculin by immunoblotting. (I).Mouse mixed brain cells were infected with HSV-2 (MOI=10) for 36 h and lysed for analysis of LDH release was measured by ELISA (J). Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗p ≤ 0.05; ∗∗p ≤ 0.01 (Mann-Whitney test, two-tailed in J).
Figure 3(A) Lysates from SH-SY5Y and THP1 cells were immunoblotted for a panel of caspases and vinculin.(B) SH-SY5Y cells were pre-treated with CASP2 inhibitor Z-VDVAD (20 μM) for 1 h and infected with HSV-2 (MOI=1). Lysates were isolated 16 hpi for immunoblot analysis.(C, SH-SY5Y cells were transfected with CASP2 gRNA-Cas9 RNPs or AAVS1 gRNA-Cas9 RNPs and infected with HSV-2. Lysates were isolated 16 h post infection and immunoblotted for GSDME, CASP2, cleaved caspase 3 (CL-CASP3), cleaved PARP (CL-PARP) and vinculin (C).G) SH-SY5Y cells were infected with HSV-2. Supernatant and lysates were isolated for identification of HMGB1 (G, 16 h).(H) SH-SY5Y cells transfected with AAVS1 or CASP2 gRNA-Cas9 RNPs and infected with HSV-2 for 16 h and examined for morphological changes by microscopy. Boxes indicate areas highlighted in the zoomed images. Arrows indicate ballooning or syncytial cells.
Figure 4(B, SH-SY5Y cells transfected with AAVS1 or BID gRNA-Cas9 RNPs were infected with HSV-2. Lysates were isolated 16 h post infection and immunoblotted for GSDME, CL-CASP3 and CL-PARP (B)C) SH-SY5Y cells transfected with AAVS1 or BID gRNA-Cas9 RNPs were infected with HSV-2. Lysates were isolated 16 h post infection and assayed for CASP3 activity (C). Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗∗p ≤ 0.001 two-way ANOVA in C(G) NADPH reductase activity was measured to evaluate mitochondria function in SH-SY5Y cells infected with HSV-2 (MOI, 1) for the indicated time interval. Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗∗p ≤ 0.001 (Mann-Whitney test, two-tailed in GSH-SY5Y cells were treated with ethidium bromide (2 μg/ml) and infected with HSV-2 (MOI=1, 16 h). Lysate were for immunoblotting for cell death markers GSDMESH-SY5Y cells were treated with ethidium bromide (2 μg/ml) and infected with HSV-2 (MOI=1, 16 h). Lysate and supernatant were for immunoblotting for cell death markers HMGB1SH-SY5Y cells were treated with ethidium bromide (2 μg/ml) and infected with HSV-2 (MOI=1, 16 h). Lysate were for immunoblotting for cell death markers GSDME SH-SY5Y cells were transfected with Poly I:C (50 μg/ml) or dsDNA (20 μg/ml) for 24 h (J).
Figure 5(A, SH-SY5Y cells were infected with HSV-2 (MOI=1) for the indicated time intervals, and immunoblotted for ER stress markers (BIP, phospho-IRE1α, phospho-eIF2α, cleaved-ATF6, CHOP)B) SH-SY5Y cells were infected with HSV-2 (MOI=1) for the indicated time intervals, and analyzed for mRNA levels of the ER stress response genes BIP, ATF6 and CHOP by RT-qPCR (B). Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001 (Mann-Whitney test, two-tailed in B(C) Lysate from SH-SY5Y cells with HSV-2 infection (MOI=1) were immunoblotted for BCL2 family proteins MCL-1, Bcl-XL, and phospho-BAD (S112). Vinculin was used as loading control.SH-SY5Y cells were pretreated with TUDCA (500 ng/ml) for 1 h and infected with HSV-2 (MOI=1). Lysates were isolated after 16 h of HSV-2 infection and subjected to immunoblotting for CL-CASP3, CL-PARP, GSDMESH-SY5Y cells were pretreated with TUDCA (500 ng/ml) for 1 h and infected with HSV-2 (MOI=1). supernatants (24 h) were analysed for LDH release (H, ELISA). Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001 two-way ANOVA in H).(L) SH-SY5Y cells transduced with lenti-plasmid BIP or empty vector control were infected with HSV-2 (MOI=1, 16 h). Lysates were isolated and immunoblotted for GSDME and tBID.
Figure 6SH-SY5Y cells were infected with HSV-2 for the indicated time intervals. Total RNA was analyzed for spliced XBP1 (XBP1s) by RT-qPCR Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001 (Mann-Whitney test, two-tailed in BSH-SY5Y cells were infected with HSV-2 for the indicated time intervals. Lysates were immunoblotted for IRE1α phosphor-JNK and XBP1s (C).(E) SH-SY5Y cells transfected with AAVS1 or IRE1A gRNA-Cas9 RNPs were infected with HSV-2 (MOI=1, 16 h). Lysates and supernatant were isolated and immunoblotted for IRE1α, GSDME, cytochrome c and HMGB1 release.G) SH-SY5Y cells were pretreated with APY29 for 1 h and following HSV-2 infection (16 h). Lysate were immunoblotted for p-JNK.(H) SH-SY5Y cells were pretreated with SP600125 (10 μM) for 1 h infected with HSV-2 (MOI=1, 16 h). Lysates and supernatant were immunoblotted for GSDME, cytochrome c, HMGB1, and vinculin.
Figure 7(B-D) iPSC-derived microglia cells were treated with the conditioned medium from HSV-2-infected control and GSDME-depleted SH-SY5Y cells for 8 h. Total RNA was isolated and analyzed for expression of IL6, TNFA, and CXCL10 by RT-qPCR. Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001 (Two-way ANOVA in B-D(E-G) Conditioned medium from SH-SY5Y cells with HSV-2 infection (MOI=1, 24 h) was transferred to THP1 cells (WT, STING-/-, MYD88-/-, TRIF-/- and MAVS-/-). Total RNA was isolated 8h later and analyzed for expression of IL6 (E), TNFA (F), and CXCL10 (G) by RT-qPCR. Data information: All data shown are representative of at least 3 independent experiments. Data are presented as mean ± s.d. in all graphs. ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001 Wilcoxon matched-pairs test in E-G).
Figure 1A NTL-2-foci localize in close proximity to mitochondria in young adult animals; hypodermis is imaged (green: NTL-2, red: TMRE (Tetramethylrhodamine, ethyl ester, perchlorate), a mitochondrial membrane potential-dependent dye) (n=3 independent experiments). B Localization of NTL-2-foci relative to mitochondria in young adult animals; body wall muscle cells are imaged (green: NTL-2, red: TOMM-20, an outer mitochondrial membrane (OMM) protein-marker of mitochondria) (n=3 independent experiments). C EDC-3-foci localize in close proximity to mitochondria in young adult animals; body wall muscle cells are imaged (green: EDC-3, red: TMRE) under control conditions (n=3 independent experiments). D Localization of EDC-3-foci relative to mitochondria in young adult animals; body wall muscle cells are imaged (red: EDC-3, green: mitochondrial matrix targeted by GFP) (n=3 independent experiments).F Immunoblot analysis of the cytoplasmic and mitochondria-containing fractions obtained from whole animal extracts, showing that NTL-2, CCF-1, EDC-3 and DCAP-2 are localized in the cytoplasm and co-precipitate with mitochondria (n=3 independent experiments).I EDC-3- and NTL-2-foci lose their specific localization close to mitochondria upon paraquat treatment of transgenic animals expressing mitochondria-targeted GFP , top: EDC-3-foci, bottom: NTL-2-foci (n=3 independent experiments).L EDC-3- and NTL-2-foci lose their specific localization in the vicinity of mitochondria upon genetic inhibition of mrps-5, top: EDC-3-foci, bottom: NTL-2-foci (n=3 independent experiments).
Figure 2A Confocal images of young adult animals expressing mitochondria-targeted GFP in body wall muscle cells upon dcap-2 or ntl-2 RNAi treatment (right panels indicate higher magnification images of inlays in left panels; right panel scale bar, 10μm). Images were acquired using a X40 objective lens (n=3 independent experiments).D Quantification of intestinal mitochondrial mass upon knockdown of either dcap-2 or ntl-2, in animals shown in Fig 2B & C. (n=3 independent experiments with at least 30 animals/experiment; ****P< 0.0001; one-way analysis of variance (ANOVA).F Representative images showing the effect of dcap-2 and ntl-2 genetic inhibition on the intestinal cell mitochondrial content in 7-day-old nematodes. (n=3 independent experiments; Images were acquired using a X5 objective lens). G Quantification of the mean GFP fluorescence intensity in the matrix of intestinal mitochondria as shown in Fig 2F (n=3 independent experiments with at least 57 animals/experiment; ****P< 0.0001; one-way analysis of variance (ANOVA)).H Quantification of the mtDNA/nDNA ratio under control conditions and upon genetic inhibition of either dcap-2 or ntl-2 (n=2 independent experiments; ***P< 0.001, *P< 0.05; one-way analysis of variance (ANOVA)).
Figure 3A Immunoblot analysis of SKN-1 protein levels upon the indicated genetic inhibitions. B Quantification showing the normalized SKN-1 protein levels (n= at least 3 independent experiments; *P<0.05, **P<0.01; Welch's one-way analysis of variance (ANOVA) followed by Dunnett's T3 multiple comparisons test).C Measurement of the promoter activity of the SKN-1 target gene gst-4 upon genetic inhibition of either dcap-2 or ntl-2 in wild-type background, daf-2 mutant background and in a mutant background (skn-1(lax120)) where SKN-1 is constitutively active (n=3 independent experiments with at least 256 animals/experiment; ****P< 0.0001; two-way analysis of variance (ANOVA), followed by Turkey's multiple comparison test).E Relative expression of skn-1 mRNA in wild-type, DCAP-2- and NTL-2- depleted animals (n=4 independent experiments, P(dcap-2(RNAi)) =0.98, P(ntl-2(RNAi))>0.99; one-way analysis of variance (ANOVA)).H Relative expression of aak-2 mRNA in wild-type, DCAP-2- and NTL-2- depleted animals (n=2 independent experiments; P(dcap-2(RNAi)) =0.85, P(ntl-2(RNAi))>0.99; one-way analysis of variance (ANOVA)).J Quantification of the mitochondrial mass in the animals shown in Fig 3I under the indicated treatments (n=3 independent experiments with at least 255 animals/experiment; *P< 0.05, **P< 0.01, ****P< 0.0001; one-way analysis of variance (ANOVA)).
Figure 4D Representative images showing an increase in NTL-2-foci upon dcap-2 genetic inhibition (n=3 independent experiments).E Quantification of NTL-2 foci upon dcap-2 genetic inhibition (n=3 independent experiments with at least 40 animals/experiment; ****P<0.0001; two-tailed unpaired t-test).F Representative images showing an increase in the signal of DCAP-2-foci upon ntl-2 genetic inhibition. Images were acquired using a X40 objective lens (n=3 independent experiments). G Respective quantification (n=3 independent experiments with at least 80 animals/experiment; ****P<0.0001; two-tailed unpaired t-test).L Quantification of the percentage of fluorescence recovery after photobleaching (FRAP) to measure the rates of de novo protein synthesis upon genetic inhibition of either ntl-2 or dcap-2 in 6-day-old-adult animals; the protein synthesis inhibitor cycloheximide (500μM) and let-363 genetic inhibition where used as controls. Measurements started after animals have recovered for 2hrs post photobleaching (n=3 independent experiments with at least 50 animals/experiment).
Figure 5A Analysis of expression of select MTPTs by real time quantitative PCR (RT-qPCR) following RNA immunoprecipitation (RIP) in anti-GFP isolates from NTL-2::GFP transgenic animals, unc-119(ed3)III (no GFP expression) and pgst-4GFP (used as an additional negative control) counterparts (n=2 independent experiments, ***P< 0.001; one-way analysis of variance (ANOVA).B Representative images showing the localization of NTL-2/storage bodies relative to mitochondria upon genetic inhibition of either tomm-20 or akap-1 (green: NTL-2, red: TMRE, a mitochondrial-membrane potential-dependent dye; n=3 independent experiments). Scale bars, 20 µm. Images were acquired using a X63 objective lens. C Quantification of the distances NTL-2/storage bodies acquire from mitochondria upon genetic inhibition of either tomm-20 or akap-1 (n=3 independent experiments with at least 45 animals/experiment; ***P< 0.001; one-way analysis of variance (ANOVA)).E Immunoblot analysis in mitochondria isolates of whole animal extracts showing the protein levels of NTL-2 contained in the isolate in control conditions and upon genetic inhibition of akap-1, tomm-20, mrps-5 and atp-3; genetic inhibition of ntl-2 is used as a control; NTL-2 protein is detected by an anti-GFP antibody in transgenic animals that contain the NTL-2::GFP protein fusion (n=2 independent experiments). F Quantification of NTL-2 protein bound to mitochondria under control conditions and upon genetic inhibitions shown in E; CTC-1 is used as a loading control for mitochondria (n=2 independent experiments; *P<0.05, **P<0.01, ***P<0.001; one-way analysis of variance (ANOVA) followed by Dunnett's test).
Figure 6A Percent survival of wild-type, DCAP-2- and NTL-2- depleted animals subjected to heat stress for 5 hrs at 37oC and then counted every 24 hours (n=4 independent experiments with at least 294 animals/experiment). B Percent survival of wild-type, DCAP-2- and NTL-2- depleted animals following paraquat (8mM) administration and counted every 24 hours (n=4 independent experiments with at least 144 animals/experiment). C Percent survival of wild-type, DCAP-2- and NTL-2- depleted animals counted every 24 hours post CCCP (15μM) treatment (n=4 independent experiments with at least 116 animals/experiment).E Knockdown of ntl-2 shortens the lifespan of wild-type animals, while NTL-2 overexpression extends lifespan.
Figure 7A Knockdown of akap-1 extends the lifespan of wild-type animals, while it does not further extend the lifespan of NTL-2 overexpressing animals. B Knockdown of akap-1 reverses the lifespan shortening of ntl-2 mutant worms. C Knockdown of akap-1 ameliorates the lifespan shortening caused by ntl-2 genetic inhibition (double RNAi administered as 1:1 mixture).
Figure 1(D-F) Box plots showing diastolic (D) and systolic (E) diameters and cardiac contractility analyses (percent fractional shortening) (F) performed using SOHA approach for controls (Hand-Gal4 and UAS-dmiR-1 sponge) and Hand>sponge dmiR-1 contexts at ages one and five weeks. n = 20 hearts. Central band corresponds to median. Whiskers correspond to Min to Max. Box corresponds to interquartile range from 25th to 75th percentile. Data information: Scale bar = 20 µm. p-value<0.05 considered statistically significant. (*) p-value = 0.033, (**) p-value =0.021, (***) p-value =0.0002. Non-parametric Mann Whitney test was performed to compare control samples and samples of interest.
Figure 2(G,G') Representative spot views generated using Imaris from in situ hybridization with miRCURY LNA probe for dmiR-1 and used for quantification of dmiR-1 levels. Spot views of dmiR-1 in hearts of one-week-old control (UAS-mblRNAi) (G) and DM1 flies (Hand>mblRNAi) (G') are shown. Each spot represents a pool of dmiR-1 transcripts detected in the same area. The zoom region in G corresponds to area in which FISH signal was quantified. Data information: Scale bar = 40 µm.(H, I) Scatter plot graph showing the signal intensity quantified in cardioblasts of one- and five-week-old flies for controls (UAS-Bru3, UAS-mblRNAi) and DM1 contexts (Hand>Bru3, Hand>mblRNAi). n = 9 hearts. Bars corresponds to mean ± SD. Data information: p-value<0.05 considered statistically significant. (*) p-value = 0.033, (**) p-value =0.021, (***) p-value =0.0002. (****) p-value<0.0001. Non-parametric Mann Whitney test was performed to compare control samples and samples of interest.